From 482a679ca8e53ac9d9d72d9733fafafedce0aded Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 5 Nov 2019 13:49:02 +0000 Subject: [PATCH 001/226] Bump version to 1.1.1dev --- .travis.yml | 2 +- Dockerfile | 4 ++-- environment.yml | 2 +- nextflow.config | 4 ++-- 4 files changed, 6 insertions(+), 6 deletions(-) diff --git a/.travis.yml b/.travis.yml index 0d477d1b..55073b54 100755 --- a/.travis.yml +++ b/.travis.yml @@ -14,7 +14,7 @@ before_install: - docker pull nfcore/atacseq:dev # Fake the tag locally so that the pipeline runs properly # Looks weird when this is :dev to :dev, but makes sense when testing code for a release (:dev to :1.0.1) - - docker tag nfcore/atacseq:dev nfcore/atacseq:1.1.0 + - docker tag nfcore/atacseq:dev nfcore/atacseq:dev install: # Install Nextflow diff --git a/Dockerfile b/Dockerfile index 7c8ffa13..eed4f3cb 100755 --- a/Dockerfile +++ b/Dockerfile @@ -7,7 +7,7 @@ COPY environment.yml / RUN conda env create -f /environment.yml && conda clean -a # Add conda installation dir to PATH (instead of doing 'conda activate') -ENV PATH /opt/conda/envs/nf-core-atacseq-1.1.0/bin:$PATH +ENV PATH /opt/conda/envs/nf-core-atacseq-1.1.1dev/bin:$PATH # Dump the details of the installed packages to a file for posterity -RUN conda env export --name nf-core-atacseq-1.1.0 > nf-core-atacseq-1.1.0.yml +RUN conda env export --name nf-core-atacseq-1.1.1dev > nf-core-atacseq-1.1.1dev.yml diff --git a/environment.yml b/environment.yml index 7291df75..5dcb2335 100755 --- a/environment.yml +++ b/environment.yml @@ -1,6 +1,6 @@ # You can use this file to create a conda environment for this pipeline: # conda env create -f environment.yml -name: nf-core-atacseq-1.1.0 +name: nf-core-atacseq-1.1.1dev channels: - conda-forge - bioconda diff --git a/nextflow.config b/nextflow.config index 0255adf4..ceface2e 100755 --- a/nextflow.config +++ b/nextflow.config @@ -92,7 +92,7 @@ params { // Container slug. Stable releases should specify release tag! // Developmental code should specify :dev -process.container = 'nfcore/atacseq:1.1.0' +process.container = 'nfcore/atacseq:dev' // Load base.config by default for all pipelines includeConfig 'conf/base.config' @@ -159,7 +159,7 @@ manifest { description = 'ATACSeq peak-calling and differential analysis pipeline.' mainScript = 'main.nf' nextflowVersion = '>=19.10.0' - version = '1.1.0' + version = '1.1.1dev' } // Function to ensure that resource requirements don't go beyond From 4cecfdd2ef23b2be91fbcdbeee7f34f1f9055eb3 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 5 Nov 2019 13:52:18 +0000 Subject: [PATCH 002/226] Update CHANGELOG --- CHANGELOG.md | 10 ++++++++++ 1 file changed, 10 insertions(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index 9abafb44..b06732c3 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -5,6 +5,16 @@ All notable changes to this project will be documented in this file. The format is based on [Keep a Changelog](http://keepachangelog.com/en/1.0.0/) and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0.html). +[Unpublished Version / DEV] + +### `Added` + +### `Fixed` + +### `Dependencies` + +### `Deprecated` + ## [1.1.0] - 2019-11-05 ### `Added` From ce452afbf4e20df43eba710057487fab300df423 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 5 Nov 2019 16:09:13 +0000 Subject: [PATCH 003/226] Fix markdownlint --- CHANGELOG.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index b06732c3..e948eed1 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -5,7 +5,7 @@ All notable changes to this project will be documented in this file. The format is based on [Keep a Changelog](http://keepachangelog.com/en/1.0.0/) and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0.html). -[Unpublished Version / DEV] +## [Unpublished Version / DEV] ### `Added` From bbfd5e5c7b0525b66b5adfe12395921510008b38 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 8 Nov 2019 21:55:47 +0000 Subject: [PATCH 004/226] Fix skip_trimming bug --- main.nf | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/main.nf b/main.nf index 1f32ea96..45b34590 100644 --- a/main.nf +++ b/main.nf @@ -521,8 +521,8 @@ process FastQC { */ if (params.skip_trimming) { ch_trimmed_reads = ch_raw_reads_trimgalore - ch_trimgalore_results_mqc = [] - ch_trimgalore_fastqc_reports_mqc = [] + ch_trimgalore_results_mqc = Channel.empty() + ch_trimgalore_fastqc_reports_mqc = Channel.empty() } else { process TrimGalore { tag "$name" From 087e00a2e909331f466d21c9b6ac73aeb38ca2aa Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 12 Nov 2019 14:04:49 +0000 Subject: [PATCH 005/226] Update CHANGELOG --- CHANGELOG.md | 2 ++ 1 file changed, 2 insertions(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index e948eed1..deb6ea8c 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -11,6 +11,8 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Fixed` +* [#118](https://github.com/nf-core/chipseq/issues/118) - Running on with SGE + ### `Dependencies` ### `Deprecated` From 2e0087bb428ca9e36dd1bb4a7c6fd468e31a3f97 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 12 Nov 2019 14:05:09 +0000 Subject: [PATCH 006/226] Remove requests --- bin/check_design.py | 1 - 1 file changed, 1 deletion(-) diff --git a/bin/check_design.py b/bin/check_design.py index 4c1436f0..d533f53e 100755 --- a/bin/check_design.py +++ b/bin/check_design.py @@ -8,7 +8,6 @@ import os import sys -import requests import argparse ############################################ From 2ab19c9e42d71665d328ea8935a83c2bf61a8f4a Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 12 Nov 2019 15:53:35 +0000 Subject: [PATCH 007/226] Fix AWS batch settings --- main.nf | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/main.nf b/main.nf index 45b34590..28f8b260 100644 --- a/main.nf +++ b/main.nf @@ -193,14 +193,14 @@ if (params.bwa_index) { /* -- AWS -- */ //////////////////////////////////////////////////// -if (workflow.profile == 'awsbatch') { +if (workflow.profile.contains('awsbatch')) { // AWSBatch sanity checking if (!params.awsqueue || !params.awsregion) exit 1, "Specify correct --awsqueue and --awsregion parameters on AWSBatch!" // Check outdir paths to be S3 buckets if running on AWSBatch // related: https://github.com/nextflow-io/nextflow/issues/813 if (!params.outdir.startsWith('s3:')) exit 1, "Outdir not on S3 - specify S3 Bucket to run on AWSBatch!" // Prevent trace files to be stored on S3 since S3 does not support rolling files. - if (workflow.tracedir.startsWith('s3:')) exit 1, "Specify a local tracedir or run without trace! S3 cannot be used for tracefiles." + if (params.tracedir.startsWith('s3:')) exit 1, "Specify a local tracedir or run without trace! S3 cannot be used for tracefiles." } /////////////////////////////////////////////////////////////////////////////// From ea48b5093f76e76cdcdb53cf858d5cc6ac46a410 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 12 Nov 2019 18:49:37 +0000 Subject: [PATCH 008/226] Fix Tpyo --- .github/PULL_REQUEST_TEMPLATE.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/.github/PULL_REQUEST_TEMPLATE.md b/.github/PULL_REQUEST_TEMPLATE.md index 2784b969..ab7fd57d 100644 --- a/.github/PULL_REQUEST_TEMPLATE.md +++ b/.github/PULL_REQUEST_TEMPLATE.md @@ -1,4 +1,4 @@ -Many thanks to contributing to nf-core/atacseq! +Many thanks for contributing to nf-core/atacseq! Please fill in the appropriate checklist below (delete whatever is not relevant). These are the most common things requested on pull requests (PRs). From e346b409e2a1c680f23bb5d4df0bdcb513df186b Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 13 Nov 2019 09:59:27 +0000 Subject: [PATCH 009/226] Change saveAlignedIntermediates to save_align_intermeds --- docs/output.md | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/docs/output.md b/docs/output.md index fe6e82a9..a9e31909 100755 --- a/docs/output.md +++ b/docs/output.md @@ -64,7 +64,7 @@ The initial QC and alignments are performed at the library-level e.g. if the sam *Output directories*: * `bwa/library/` - The files resulting from the alignment of individual libraries are not saved by default so this directory will not be present in your results. You can override this behaviour with the use of the `--saveAlignedIntermediates` flag in which case it will contain the coordinate sorted alignment files in [`*.bam`](https://samtools.github.io/hts-specs/SAMv1.pdf) format. + The files resulting from the alignment of individual libraries are not saved by default so this directory will not be present in your results. You can override this behaviour with the use of the `--save_align_intermeds` flag in which case it will contain the coordinate sorted alignment files in [`*.bam`](https://samtools.github.io/hts-specs/SAMv1.pdf) format. * `bwa/library/samtools_stats/` SAMtools `*.flagstat`, `*.idxstats` and `*.stats` files generated from the alignment files. @@ -100,7 +100,7 @@ The library-level alignments associated with the same sample are merged and subs *Output directories*: * `bwa/mergedLibrary/` - Merged library-level, coordinate sorted `*.bam` files after the marking of duplicates, and filtering based on various criteria. The file suffix for the final filtered files will be `*.mLb.clN.*`. If you specify the `--saveAlignedIntermediates` parameter then two additional sets of files will be present. These represent the unfiltered alignments with duplicates marked (`*.mLb.mkD.*`), and in the case of paired-end datasets the filtered alignments before the removal of orphan read pairs (`*.mLb.flT.*`). + Merged library-level, coordinate sorted `*.bam` files after the marking of duplicates, and filtering based on various criteria. The file suffix for the final filtered files will be `*.mLb.clN.*`. If you specify the `--save_align_intermeds` parameter then two additional sets of files will be present. These represent the unfiltered alignments with duplicates marked (`*.mLb.mkD.*`), and in the case of paired-end datasets the filtered alignments before the removal of orphan read pairs (`*.mLb.flT.*`). * `bwa/mergedLibrary/samtools_stats/` SAMtools `*.flagstat`, `*.idxstats` and `*.stats` files generated from the alignment files. * `bwa/mergedLibrary/picard_metrics/` From e377f738d101561bc10445d713decdae1c4aafcd Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 13 Nov 2019 10:21:44 +0000 Subject: [PATCH 010/226] Change skipMergeReplicate to skip_merge_replicates --- docs/output.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/docs/output.md b/docs/output.md index a9e31909..c4c2f6fc 100755 --- a/docs/output.md +++ b/docs/output.md @@ -259,7 +259,7 @@ The alignments associated with all of the replicates from the same experimental File names in the resulting directory (i.e. `bwa/mergedReplicate/`) will have the '`.mRp.`' suffix to denote **m**erged **R**e**p**licates. -You can skip this portion of the analysis by specifying the `--skipMergeReplicate` parameter. +You can skip this portion of the analysis by specifying the `--skip_merge_replicates` parameter. >NB: Merged library-level alignments will be used for read counting relative to the consensus merged replicate-level peakset. This is the only way in which differential analysis can be performed at the merged replicate-level. From 7fc74257c688034e29ed1b63358c9a176995aaef Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 14 Nov 2019 12:19:12 +0000 Subject: [PATCH 011/226] Make script work with Python3 --- bin/check_design.py | 14 +++++++------- 1 file changed, 7 insertions(+), 7 deletions(-) diff --git a/bin/check_design.py b/bin/check_design.py index d533f53e..145ef909 100755 --- a/bin/check_design.py +++ b/bin/check_design.py @@ -41,7 +41,7 @@ def check_design(DesignFileIn,DesignFileOut): fin = open(DesignFileIn,'r') header = fin.readline().strip().split(',') if header != HEADER: - print "{} header: {} != {}".format(ERROR_STR,','.join(header),','.join(HEADER)) + print("{} header: {} != {}".format(ERROR_STR,','.join(header),','.join(HEADER))) sys.exit(1) numColList = [] @@ -54,25 +54,25 @@ def check_design(DesignFileIn,DesignFileOut): ## CHECK VALID NUMBER OF COLUMNS PER SAMPLE numCols = len(lspl) if numCols not in [3,4]: - print "{}: Invalid number of columns (3 for single-end or 4 for paired-end)!\nLine: '{}'".format(ERROR_STR,line.strip()) + print("{}: Invalid number of columns (3 for single-end or 4 for paired-end)!\nLine: '{}'".format(ERROR_STR,line.strip())) sys.exit(1) numColList.append(numCols) ## CHECK GROUP COLUMN HAS NO SPACES group,replicate,fastQFiles = lspl[0],lspl[1],lspl[2:] if group.find(' ') != -1: - print "{}: Group id contains spaces!\nLine: '{}'".format(ERROR_STR,line.strip()) + print("{}: Group id contains spaces!\nLine: '{}'".format(ERROR_STR,line.strip())) sys.exit(1) ## CHECK REPLICATE COLUMN IS INTEGER if not replicate.isdigit(): - print "{}: Replicate id not an integer!\nLine: '{}'".format(ERROR_STR,line.strip()) + print("{}: Replicate id not an integer!\nLine: '{}'".format(ERROR_STR,line.strip())) sys.exit(1) for fastq in fastQFiles: ## CHECK FASTQ FILE EXTENSION if fastq[-9:] != '.fastq.gz' and fastq[-6:] != '.fq.gz': - print "{}: FastQ file has incorrect extension (has to be '.fastq.gz' or 'fq.gz') - {}\nLine: '{}'".format(ERROR_STR,fastq,line.strip()) + print("{}: FastQ file has incorrect extension (has to be '.fastq.gz' or 'fq.gz') - {}\nLine: '{}'".format(ERROR_STR,fastq,line.strip())) sys.exit(1) ## CREATE GROUP MAPPING DICT = {GROUP_ID: {REPLICATE_ID:[[FASTQ_FILES]]} @@ -89,7 +89,7 @@ def check_design(DesignFileIn,DesignFileOut): ## CHECK IF DATA IS PAIRED-END OR SINGLE-END AND NOT A MIXTURE if min(numColList) != max(numColList): - print "{}: Mixture of paired-end and single-end reads!".format(ERROR_STR) + print("{}: Mixture of paired-end and single-end reads!".format(ERROR_STR)) sys.exit(1) ## CHECK IF MULTIPLE GROUPS EXIST @@ -106,7 +106,7 @@ def check_design(DesignFileIn,DesignFileOut): ## CHECK THAT REPLICATE IDS ARE IN FORMAT 1.. uniq_rep_ids = set(groupRepDict[group].keys()) if len(uniq_rep_ids) != max(uniq_rep_ids): - print "{}: Replicate IDs must start with 1..\nGroup: {}, Replicate IDs: {}".format(ERROR_STR,group,list(uniq_rep_ids)) + print("{}: Replicate IDs must start with 1..\nGroup: {}, Replicate IDs: {}".format(ERROR_STR,group,list(uniq_rep_ids))) sys.exit(1) numRepList.append(max(uniq_rep_ids)) From 449157a60cfaaa7bf3826c0fc49e40d7aa888e14 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 14 Nov 2019 12:19:19 +0000 Subject: [PATCH 012/226] Make script work with Python3 --- bin/scrape_software_versions.py | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/bin/scrape_software_versions.py b/bin/scrape_software_versions.py index 0b76c7a6..022a8758 100755 --- a/bin/scrape_software_versions.py +++ b/bin/scrape_software_versions.py @@ -54,9 +54,9 @@ results[k] = "v{}".format(match.group(1)) except IOError: results[k] = False - + # Remove software set to false in results -for k in results: +for k in list(results): if not results[k]: del(results[k]) From 0e4cd20480ff9070a49cf58b7e6fde9ca8b3b279 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 14 Nov 2019 13:29:16 +0000 Subject: [PATCH 013/226] Update software to latest in environment --- environment.yml | 39 ++++++++++++++++++++------------------- 1 file changed, 20 insertions(+), 19 deletions(-) diff --git a/environment.yml b/environment.yml index 5dcb2335..33741853 100755 --- a/environment.yml +++ b/environment.yml @@ -7,34 +7,35 @@ channels: - defaults dependencies: ## conda-forge packages - - gawk=4.2.1 - - r-base=3.4.1 - - r-optparse=1.6.0 + - gawk=5.0.1 + - r-base=3.6.1 + - r-optparse=1.6.4 - r-rcolorbrewer=1.1_2 - - r-ggplot2=3.1.0 + - r-ggplot2=3.2.1 - r-reshape2=1.4.3 - r-scales=1.0.0 - - r-pheatmap=1.0.10 - - r-lattice=0.20_35 - - r-upsetr=1.3.3 - - r-xfun=0.3 + - r-pheatmap=1.0.12 + - r-lattice=0.20_38 + - r-upsetr=1.4.0 + - r-xfun=0.11 ## bioconda packages - fastqc=0.11.8 - - trim-galore=0.5.0 + - trim-galore=0.6.4 - bwa=0.7.17 - samtools=1.9 - - picard=2.19.0 + - picard=2.21.3 - bamtools=2.5.1 - - pysam=0.15.2 - - bedtools=2.27.1 - - preseq=2.0.3 - - deeptools=3.2.1 - - ucsc-bedgraphtobigwig=377 - - macs2=2.1.2 - - homer=4.9.1 + - pysam=0.15.3 + - bedtools=2.29.0 + - ucsc-bedgraphtobigwig=357 + - deeptools=3.3.1 + - macs2=2.2.5 + - genrich=0.6 + - homer=4.10 - ataqv=1.0.0 - subread=1.6.4 + - preseq=2.0.3 - multiqc=1.7 - - bioconductor-deseq2=1.20.0 - - bioconductor-vsn=3.46.0 + - bioconductor-deseq2=1.26.0 + - bioconductor-vsn=3.54.0 From b05eb47bb779103c512d04de5d0f3e5e70d5d18a Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 14 Nov 2019 13:53:23 +0000 Subject: [PATCH 014/226] Update CHANGELOG --- CHANGELOG.md | 21 +++++++++++++++++++++ 1 file changed, 21 insertions(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index deb6ea8c..3f003c04 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -12,9 +12,30 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Fixed` * [#118](https://github.com/nf-core/chipseq/issues/118) - Running on with SGE +* Make executables in `bin/` compatible with Python 3 ### `Dependencies` +* Add genrich `0.6` +* Update gawk `4.2.1` -> `5.0.1` +* Update r-base `3.4.1` -> `3.6.1` +* Update r-optparse `1.6.0` -> `1.6.4` +* Update r-ggplot2 `3.1.0` -> `3.2.1` +* Update r-pheatmap `1.0.10` -> `1.0.12` +* Update r-lattice `0.20_35` -> `0.20_38` +* Update r-upsetr `1.3.3` -> `1.4.0` +* Update r-xfun `0.3` -> `0.11` +* Update trim-galore `0.5.0` -> `0.6.4` +* Update picard `2.19.0` -> `2.21.3` +* Update pysam `0.15.2` -> `0.15.3` +* Update bedtools `2.27.1` -> `2.29.0` +* Update ucsc-bedgraphtobigwig `377` -> `357` +* Update deeptools `3.2.1` -> `3.3.1` +* Update macs2 `2.1.2` -> `2.2.5` +* Update homer `4.9.1` -> `4.10` +* Update bioconductor-deseq2 `1.20.0` -> `1.26.0` +* Update bioconductor-vsn `3.46.0` -> `3.54.0` + ### `Deprecated` ## [1.1.0] - 2019-11-05 From 3f103ab99021dcce7881bd50cca79308b8ed8b99 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 14 Nov 2019 14:32:44 +0000 Subject: [PATCH 015/226] Replace has_key with in --- bin/check_design.py | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/bin/check_design.py b/bin/check_design.py index 145ef909..d55e2cec 100755 --- a/bin/check_design.py +++ b/bin/check_design.py @@ -77,9 +77,9 @@ def check_design(DesignFileIn,DesignFileOut): ## CREATE GROUP MAPPING DICT = {GROUP_ID: {REPLICATE_ID:[[FASTQ_FILES]]} replicate = int(replicate) - if not groupRepDict.has_key(group): + if group not in groupRepDict: groupRepDict[group] = {} - if not groupRepDict[group].has_key(replicate): + if replicate not in groupRepDict[group]: groupRepDict[group][replicate] = [] groupRepDict[group][replicate].append(fastQFiles) From 072689dc37172542242ab2136abe4132fcdfdbc6 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 14 Nov 2019 14:32:51 +0000 Subject: [PATCH 016/226] Replace has_key with in --- bin/macs2_merged_expand.py | 18 +++++++++--------- 1 file changed, 9 insertions(+), 9 deletions(-) diff --git a/bin/macs2_merged_expand.py b/bin/macs2_merged_expand.py index 10889947..55c3a057 100755 --- a/bin/macs2_merged_expand.py +++ b/bin/macs2_merged_expand.py @@ -87,9 +87,9 @@ def macs2_merged_expand(MergedIntervalTxtFile,SampleNameList,OutFile,isNarrow=Fa groupDict = {} for sID in ['_'.join(x.split('_')[:-2]) for x in names]: gID = '_'.join(sID.split('_')[:-1]) - if not groupDict.has_key(gID): + if gID not in groupDict: groupDict[gID] = [] - if not sID in groupDict[gID]: + if sID not in groupDict[gID]: groupDict[gID].append(sID) ## GET SAMPLES THAT PASS REPLICATE THRESHOLD @@ -103,23 +103,23 @@ def macs2_merged_expand(MergedIntervalTxtFile,SampleNameList,OutFile,isNarrow=Fa for idx in range(len(names)): sample = '_'.join(names[idx].split('_')[:-2]) if sample in passRepThreshList: - if not fcDict.has_key(sample): + if sample not in fcDict: fcDict[sample] = [] fcDict[sample].append(str(fcs[idx])) - if not qvalDict.has_key(sample): + if sample not in qvalDict: qvalDict[sample] = [] qvalDict[sample].append(str(qvals[idx])) - if not pvalDict.has_key(sample): + if sample not in pvalDict: pvalDict[sample] = [] pvalDict[sample].append(str(pvals[idx])) - if not startDict.has_key(sample): + if sample not in startDict: startDict[sample] = [] startDict[sample].append(str(starts[idx])) - if not endDict.has_key(sample): + if sample not in endDict: endDict[sample] = [] endDict[sample].append(str(ends[idx])) if isNarrow: - if not summitDict.has_key(sample): + if sample not in summitDict: summitDict[sample] = [] summitDict[sample].append(str(summits[idx])) @@ -138,7 +138,7 @@ def macs2_merged_expand(MergedIntervalTxtFile,SampleNameList,OutFile,isNarrow=Fa fout.write('\t'.join(oList) + '\n') tsamples = tuple(sorted(samples)) - if not combFreqDict.has_key(tsamples): + if tsamples not in combFreqDict: combFreqDict[tsamples] = 0 combFreqDict[tsamples] += 1 totalOutIntervals += 1 From 9838c60b9d555815c36912515375055853591238 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 14 Nov 2019 14:58:20 +0000 Subject: [PATCH 017/226] Pin pysam version --- environment.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/environment.yml b/environment.yml index 33741853..09d992c6 100755 --- a/environment.yml +++ b/environment.yml @@ -26,7 +26,7 @@ dependencies: - samtools=1.9 - picard=2.21.3 - bamtools=2.5.1 - - pysam=0.15.3 + - pysam=0.15.2 ## Keep this version for bampe_rm_orphan.py - bedtools=2.29.0 - ucsc-bedgraphtobigwig=357 - deeptools=3.3.1 From 33c09f970ab78f37ac6f0f039432922a16923bd5 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 14 Nov 2019 16:19:41 +0000 Subject: [PATCH 018/226] Update pysam again --- environment.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/environment.yml b/environment.yml index 09d992c6..33741853 100755 --- a/environment.yml +++ b/environment.yml @@ -26,7 +26,7 @@ dependencies: - samtools=1.9 - picard=2.21.3 - bamtools=2.5.1 - - pysam=0.15.2 ## Keep this version for bampe_rm_orphan.py + - pysam=0.15.3 - bedtools=2.29.0 - ucsc-bedgraphtobigwig=357 - deeptools=3.3.1 From 25b2fc408480ee8abd2c051c35438c36128dc267 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 14 Nov 2019 16:47:19 +0000 Subject: [PATCH 019/226] Update pysam calls --- bin/bampe_rm_orphan.py | 11 ++++++----- 1 file changed, 6 insertions(+), 5 deletions(-) diff --git a/bin/bampe_rm_orphan.py b/bin/bampe_rm_orphan.py index c11d4e67..97f2fc2a 100755 --- a/bin/bampe_rm_orphan.py +++ b/bin/bampe_rm_orphan.py @@ -61,10 +61,11 @@ def bampe_rm_orphan(BAMIn,BAMOut,onlyFRPairs=False): ## ITERATE THROUGH BAM FILE EOF = 0 - SAMFin = pysam.Samfile(BAMIn,"rb") - SAMFout = pysam.Samfile(BAMOut, "wb",header=SAMFin.header) - currRead = SAMFin.next() - for read in SAMFin: + SAMFin = pysam.AlignmentFile(BAMIn, "rb") + SAMFout = pysam.AlignmentFile(BAMOut, "wb", header=SAMFin.header) + iter = SAMFin.fetch(until_eof=True) + currRead = iter.__next__() + for read in iter: totalReads += 1 if currRead.qname == read.qname: pair1 = currRead; pair2 = read @@ -103,7 +104,7 @@ def bampe_rm_orphan(BAMIn,BAMOut,onlyFRPairs=False): ## RESET COUNTER try: totalReads += 1 - currRead = SAMFin.next() + currRead = iter.__next__() except: StopIteration EOF = 1 From e747a96d79bfa952cade88943222e351bdc32937 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 14 Nov 2019 16:59:59 +0000 Subject: [PATCH 020/226] Update CHANGELOG --- CHANGELOG.md | 1 + 1 file changed, 1 insertion(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index 3f003c04..99b2793f 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -17,6 +17,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Dependencies` * Add genrich `0.6` +* Add r-stringi `1.4.3` * Update gawk `4.2.1` -> `5.0.1` * Update r-base `3.4.1` -> `3.6.1` * Update r-optparse `1.6.0` -> `1.6.4` From 572aed56a1d24bee0f9810b7efd673f8ce27db06 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 14 Nov 2019 17:00:05 +0000 Subject: [PATCH 021/226] Add r-stringi 1.4.3 --- environment.yml | 1 + 1 file changed, 1 insertion(+) diff --git a/environment.yml b/environment.yml index 33741853..b30d99bb 100755 --- a/environment.yml +++ b/environment.yml @@ -18,6 +18,7 @@ dependencies: - r-lattice=0.20_38 - r-upsetr=1.4.0 - r-xfun=0.11 + - r-stringi=1.4.3 ## bioconda packages - fastqc=0.11.8 From b5cd1264c31f1509eb09fa6da5ca0feaae944d49 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 14 Nov 2019 17:45:47 +0000 Subject: [PATCH 022/226] Add channel --- environment.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/environment.yml b/environment.yml index b30d99bb..04788280 100755 --- a/environment.yml +++ b/environment.yml @@ -18,7 +18,7 @@ dependencies: - r-lattice=0.20_38 - r-upsetr=1.4.0 - r-xfun=0.11 - - r-stringi=1.4.3 + - r::r-stringi=1.4.3 ## bioconda packages - fastqc=0.11.8 From c352ebdcfa3986a3ee4f33ecc04ad428b64f6a7e Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 14 Nov 2019 17:50:40 +0000 Subject: [PATCH 023/226] Use next() instead of __next__ --- bin/bampe_rm_orphan.py | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/bin/bampe_rm_orphan.py b/bin/bampe_rm_orphan.py index 97f2fc2a..439a53ee 100755 --- a/bin/bampe_rm_orphan.py +++ b/bin/bampe_rm_orphan.py @@ -64,7 +64,7 @@ def bampe_rm_orphan(BAMIn,BAMOut,onlyFRPairs=False): SAMFin = pysam.AlignmentFile(BAMIn, "rb") SAMFout = pysam.AlignmentFile(BAMOut, "wb", header=SAMFin.header) iter = SAMFin.fetch(until_eof=True) - currRead = iter.__next__() + currRead = next(iter) for read in iter: totalReads += 1 if currRead.qname == read.qname: @@ -104,7 +104,7 @@ def bampe_rm_orphan(BAMIn,BAMOut,onlyFRPairs=False): ## RESET COUNTER try: totalReads += 1 - currRead = iter.__next__() + currRead = next(iter) except: StopIteration EOF = 1 From ee6fd720b07be94232b167a616a2427edcde5358 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 14 Nov 2019 18:49:54 +0000 Subject: [PATCH 024/226] skip_plot_profile to bypass MultiQC error --- conf/test.config | 2 ++ 1 file changed, 2 insertions(+) diff --git a/conf/test.config b/conf/test.config index 0435d184..bd55970e 100755 --- a/conf/test.config +++ b/conf/test.config @@ -28,4 +28,6 @@ params { // For speed to avoid CI time-out fingerprint_bins = 100 + + skip_plot_profile = true } From 1938bad75551e17c05efca850e6cc982fc3c44b2 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 14 Nov 2019 23:41:51 +0000 Subject: [PATCH 025/226] Add link to MultiQC issue --- conf/test.config | 2 ++ 1 file changed, 2 insertions(+) diff --git a/conf/test.config b/conf/test.config index bd55970e..9b712466 100755 --- a/conf/test.config +++ b/conf/test.config @@ -29,5 +29,7 @@ params { // For speed to avoid CI time-out fingerprint_bins = 100 + // MultiQC v1.7 bug with Python3 + // See https://github.com/ewels/MultiQC/issues/1057 skip_plot_profile = true } From 2f38051136cf2a438cce86c8970a7d657a5b9ed3 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 14 Nov 2019 23:58:44 +0000 Subject: [PATCH 026/226] Finalllyyyyyyyyy working --- environment.yml | 60 ++++++++++++++++++++++++------------------------- 1 file changed, 30 insertions(+), 30 deletions(-) diff --git a/environment.yml b/environment.yml index 04788280..2139ea1a 100755 --- a/environment.yml +++ b/environment.yml @@ -7,36 +7,36 @@ channels: - defaults dependencies: ## conda-forge packages - - gawk=5.0.1 - - r-base=3.6.1 - - r-optparse=1.6.4 - - r-rcolorbrewer=1.1_2 - - r-ggplot2=3.2.1 - - r-reshape2=1.4.3 - - r-scales=1.0.0 - - r-pheatmap=1.0.12 - - r-lattice=0.20_38 - - r-upsetr=1.4.0 - - r-xfun=0.11 + - conda-forge::gawk=5.0.1 + - conda-forge::r-base=3.6.1 + - conda-forge::r-optparse=1.6.4 + - conda-forge::r-rcolorbrewer=1.1_2 + - conda-forge::r-ggplot2=3.2.1 + - conda-forge::r-reshape2=1.4.3 + - conda-forge::r-scales=1.0.0 + - conda-forge::r-pheatmap=1.0.12 + - conda-forge::r-lattice=0.20_38 + - conda-forge::r-upsetr=1.4.0 + - conda-forge::r-xfun=0.11 - r::r-stringi=1.4.3 ## bioconda packages - - fastqc=0.11.8 - - trim-galore=0.6.4 - - bwa=0.7.17 - - samtools=1.9 - - picard=2.21.3 - - bamtools=2.5.1 - - pysam=0.15.3 - - bedtools=2.29.0 - - ucsc-bedgraphtobigwig=357 - - deeptools=3.3.1 - - macs2=2.2.5 - - genrich=0.6 - - homer=4.10 - - ataqv=1.0.0 - - subread=1.6.4 - - preseq=2.0.3 - - multiqc=1.7 - - bioconductor-deseq2=1.26.0 - - bioconductor-vsn=3.54.0 + - bioconda::fastqc=0.11.8 + - bioconda::trim-galore=0.6.4 + - bioconda::bwa=0.7.17 + - bioconda::samtools=1.9 + - bioconda::picard=2.21.3 + - bioconda::bamtools=2.5.1 + - bioconda::pysam=0.15.3 + - bioconda::bedtools=2.29.0 + - bioconda::ucsc-bedgraphtobigwig=357 + - bioconda::deeptools=3.3.1 + - bioconda::macs2=2.2.5 + - bioconda::genrich=0.6 + - bioconda::homer=4.10 + - bioconda::ataqv=1.0.0 + - bioconda::subread=1.6.4 + - bioconda::preseq=2.0.3 + - bioconda::multiqc=1.7 + - bioconda::bioconductor-deseq2=1.26.0 + - bioconda::bioconductor-vsn=3.54.0 From bb42deec21f5b50da1f823559a616a3a6b3e037e Mon Sep 17 00:00:00 2001 From: Drew Behrens Date: Thu, 21 Nov 2019 15:08:54 +0100 Subject: [PATCH 027/226] updated TrimGalore for mult-core support --- CHANGELOG.md | 2 ++ environment.yml | 1 + main.nf | 6 +++--- 3 files changed, 6 insertions(+), 3 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 99b2793f..45c1b31a 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -13,6 +13,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * [#118](https://github.com/nf-core/chipseq/issues/118) - Running on with SGE * Make executables in `bin/` compatible with Python 3 +* [#63](https://github.com/nf-core/atacseq/issues/63) - Added multicore support for Trim Galore! ### `Dependencies` @@ -36,6 +37,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * Update homer `4.9.1` -> `4.10` * Update bioconductor-deseq2 `1.20.0` -> `1.26.0` * Update bioconductor-vsn `3.46.0` -> `3.54.0` +* Added pigz `2.3.4` ### `Deprecated` diff --git a/environment.yml b/environment.yml index 2139ea1a..65563398 100755 --- a/environment.yml +++ b/environment.yml @@ -18,6 +18,7 @@ dependencies: - conda-forge::r-lattice=0.20_38 - conda-forge::r-upsetr=1.4.0 - conda-forge::r-xfun=0.11 + - conda-forge::pigz=2.3.4 - r::r-stringi=1.4.3 ## bioconda packages diff --git a/main.nf b/main.nf index 28f8b260..6673bc9e 100644 --- a/main.nf +++ b/main.nf @@ -526,7 +526,7 @@ if (params.skip_trimming) { } else { process TrimGalore { tag "$name" - label 'process_long' + label 'process_medium' publishDir "${params.outdir}/trim_galore", mode: 'copy', saveAs: { filename -> if (filename.endsWith(".html")) "fastqc/$filename" @@ -553,13 +553,13 @@ if (params.skip_trimming) { if (params.single_end) { """ [ ! -f ${name}.fastq.gz ] && ln -s $reads ${name}.fastq.gz - trim_galore --fastqc --gzip $c_r1 $tpc_r1 $nextseq ${name}.fastq.gz + trim_galore -j $task.cpus --fastqc --gzip $c_r1 $tpc_r1 $nextseq ${name}.fastq.gz """ } else { """ [ ! -f ${name}_1.fastq.gz ] && ln -s ${reads[0]} ${name}_1.fastq.gz [ ! -f ${name}_2.fastq.gz ] && ln -s ${reads[1]} ${name}_2.fastq.gz - trim_galore --paired --fastqc --gzip $c_r1 $c_r2 $tpc_r1 $tpc_r2 $nextseq ${name}_1.fastq.gz ${name}_2.fastq.gz + trim_galore -j $task.cpus --paired --fastqc --gzip $c_r1 $c_r2 $tpc_r1 $tpc_r2 $nextseq ${name}_1.fastq.gz ${name}_2.fastq.gz """ } } From 32c7afa74564bed1899e8d5b9aff461dafb9e98c Mon Sep 17 00:00:00 2001 From: Drew Behrens Date: Thu, 21 Nov 2019 15:56:08 +0100 Subject: [PATCH 028/226] Update CHANGELOG.md Co-Authored-By: Harshil Patel --- CHANGELOG.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 45c1b31a..42297b6f 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -37,7 +37,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * Update homer `4.9.1` -> `4.10` * Update bioconductor-deseq2 `1.20.0` -> `1.26.0` * Update bioconductor-vsn `3.46.0` -> `3.54.0` -* Added pigz `2.3.4` +* Add pigz `2.3.4` ### `Deprecated` From 593233aac3fcf56e525aa48dde46376f44ed5c65 Mon Sep 17 00:00:00 2001 From: Drew Behrens Date: Thu, 21 Nov 2019 15:56:20 +0100 Subject: [PATCH 029/226] Update main.nf Co-Authored-By: Harshil Patel --- main.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/main.nf b/main.nf index 6673bc9e..ad67b7ec 100644 --- a/main.nf +++ b/main.nf @@ -553,7 +553,7 @@ if (params.skip_trimming) { if (params.single_end) { """ [ ! -f ${name}.fastq.gz ] && ln -s $reads ${name}.fastq.gz - trim_galore -j $task.cpus --fastqc --gzip $c_r1 $tpc_r1 $nextseq ${name}.fastq.gz + trim_galore --cores $task.cpus --fastqc --gzip $c_r1 $tpc_r1 $nextseq ${name}.fastq.gz """ } else { """ From fbdef55bf6d2a304e2c33d8da155dff66d6bb53b Mon Sep 17 00:00:00 2001 From: Drew Behrens Date: Thu, 21 Nov 2019 15:56:44 +0100 Subject: [PATCH 030/226] Apply suggestions from code review Co-Authored-By: Harshil Patel --- main.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/main.nf b/main.nf index ad67b7ec..816fead7 100644 --- a/main.nf +++ b/main.nf @@ -559,7 +559,7 @@ if (params.skip_trimming) { """ [ ! -f ${name}_1.fastq.gz ] && ln -s ${reads[0]} ${name}_1.fastq.gz [ ! -f ${name}_2.fastq.gz ] && ln -s ${reads[1]} ${name}_2.fastq.gz - trim_galore -j $task.cpus --paired --fastqc --gzip $c_r1 $c_r2 $tpc_r1 $tpc_r2 $nextseq ${name}_1.fastq.gz ${name}_2.fastq.gz + trim_galore --cores $task.cpus --paired --fastqc --gzip $c_r1 $c_r2 $tpc_r1 $tpc_r2 $nextseq ${name}_1.fastq.gz ${name}_2.fastq.gz """ } } From 3b77b99e2dc533a2b66a121d7c54c9c961c22247 Mon Sep 17 00:00:00 2001 From: Drew Behrens Date: Thu, 21 Nov 2019 16:02:26 +0100 Subject: [PATCH 031/226] Update CHANGELOG.md --- CHANGELOG.md | 3 ++- 1 file changed, 2 insertions(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 42297b6f..94b3268f 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -9,11 +9,12 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Added` +* [#63](https://github.com/nf-core/atacseq/issues/63) - Added multicore support for Trim Galore! + ### `Fixed` * [#118](https://github.com/nf-core/chipseq/issues/118) - Running on with SGE * Make executables in `bin/` compatible with Python 3 -* [#63](https://github.com/nf-core/atacseq/issues/63) - Added multicore support for Trim Galore! ### `Dependencies` From 78b15f2e35f3c078199aa02a693b19296cb51388 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sun, 24 Nov 2019 16:09:17 +0000 Subject: [PATCH 032/226] Update CHANGELOG --- CHANGELOG.md | 3 ++- 1 file changed, 2 insertions(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 94b3268f..f11055d4 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -20,6 +20,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * Add genrich `0.6` * Add r-stringi `1.4.3` +* Add pigz `2.3.4` * Update gawk `4.2.1` -> `5.0.1` * Update r-base `3.4.1` -> `3.6.1` * Update r-optparse `1.6.0` -> `1.6.4` @@ -36,9 +37,9 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * Update deeptools `3.2.1` -> `3.3.1` * Update macs2 `2.1.2` -> `2.2.5` * Update homer `4.9.1` -> `4.10` +* Update multiqc `1.7` -> `1.8` * Update bioconductor-deseq2 `1.20.0` -> `1.26.0` * Update bioconductor-vsn `3.46.0` -> `3.54.0` -* Add pigz `2.3.4` ### `Deprecated` From f62c4b3652346477a1f9b4f815b410d7c0b5051d Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sun, 24 Nov 2019 16:09:27 +0000 Subject: [PATCH 033/226] Update MultiQC to v1.8 --- environment.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/environment.yml b/environment.yml index 65563398..7c80e4ed 100755 --- a/environment.yml +++ b/environment.yml @@ -38,6 +38,6 @@ dependencies: - bioconda::ataqv=1.0.0 - bioconda::subread=1.6.4 - bioconda::preseq=2.0.3 - - bioconda::multiqc=1.7 + - bioconda::multiqc=1.8 - bioconda::bioconductor-deseq2=1.26.0 - bioconda::bioconductor-vsn=3.54.0 From 197ca51992ab22b0c1189557a2f2479e3c50bc8a Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sun, 24 Nov 2019 16:16:18 +0000 Subject: [PATCH 034/226] Update CHANGELOG --- CHANGELOG.md | 3 ++- 1 file changed, 2 insertions(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index f11055d4..fc18f302 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -28,8 +28,9 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * Update r-pheatmap `1.0.10` -> `1.0.12` * Update r-lattice `0.20_35` -> `0.20_38` * Update r-upsetr `1.3.3` -> `1.4.0` +* Update r-scales `1.0.0` -> `1.1.0` * Update r-xfun `0.3` -> `0.11` -* Update trim-galore `0.5.0` -> `0.6.4` +* Update trim-galore `0.5.0` -> `0.6.5` * Update picard `2.19.0` -> `2.21.3` * Update pysam `0.15.2` -> `0.15.3` * Update bedtools `2.27.1` -> `2.29.0` From fa4727d838c3cfa0abc2a06de0be4393e984f6b8 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sun, 24 Nov 2019 16:16:30 +0000 Subject: [PATCH 035/226] Update misc software --- environment.yml | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/environment.yml b/environment.yml index 7c80e4ed..b89ccc24 100755 --- a/environment.yml +++ b/environment.yml @@ -13,7 +13,7 @@ dependencies: - conda-forge::r-rcolorbrewer=1.1_2 - conda-forge::r-ggplot2=3.2.1 - conda-forge::r-reshape2=1.4.3 - - conda-forge::r-scales=1.0.0 + - conda-forge::r-scales=1.1.0 - conda-forge::r-pheatmap=1.0.12 - conda-forge::r-lattice=0.20_38 - conda-forge::r-upsetr=1.4.0 @@ -23,7 +23,7 @@ dependencies: ## bioconda packages - bioconda::fastqc=0.11.8 - - bioconda::trim-galore=0.6.4 + - bioconda::trim-galore=0.6.5 - bioconda::bwa=0.7.17 - bioconda::samtools=1.9 - bioconda::picard=2.21.3 From 8aa1cb0409872799741ac22e979f54dcc3a216ed Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sun, 24 Nov 2019 16:19:00 +0000 Subject: [PATCH 036/226] Update TrimGalore logic --- main.nf | 20 +++++++++++++++++--- 1 file changed, 17 insertions(+), 3 deletions(-) diff --git a/main.nf b/main.nf index 816fead7..45c0e2bc 100644 --- a/main.nf +++ b/main.nf @@ -526,7 +526,7 @@ if (params.skip_trimming) { } else { process TrimGalore { tag "$name" - label 'process_medium' + label 'process_high' publishDir "${params.outdir}/trim_galore", mode: 'copy', saveAs: { filename -> if (filename.endsWith(".html")) "fastqc/$filename" @@ -544,22 +544,36 @@ if (params.skip_trimming) { file "*.{zip,html}" into ch_trimgalore_fastqc_reports_mqc script: + // Calculate number of --cores for TrimGalore based on value of task.cpus + // See: https://github.com/FelixKrueger/TrimGalore/blob/master/Changelog.md#version-060-release-on-1-mar-2019 + // See: https://github.com/nf-core/atacseq/pull/65#issuecomment-557839116 + def cores = 1 + if (task.cpus) { + tcores = (((task.cpus as int) - 3) / 3) as int + if (tcores > 1) { + cores = tcores + } + } + + // Added soft-links to original fastqs for consistent naming in MultiQC c_r1 = params.clip_r1 > 0 ? "--clip_r1 ${params.clip_r1}" : '' c_r2 = params.clip_r2 > 0 ? "--clip_r2 ${params.clip_r2}" : '' tpc_r1 = params.three_prime_clip_r1 > 0 ? "--three_prime_clip_r1 ${params.three_prime_clip_r1}" : '' tpc_r2 = params.three_prime_clip_r2 > 0 ? "--three_prime_clip_r2 ${params.three_prime_clip_r2}" : '' nextseq = params.trim_nextseq > 0 ? "--nextseq ${params.trim_nextseq}" : '' + + // Added soft-links to original fastqs for consistent naming in MultiQC if (params.single_end) { """ [ ! -f ${name}.fastq.gz ] && ln -s $reads ${name}.fastq.gz - trim_galore --cores $task.cpus --fastqc --gzip $c_r1 $tpc_r1 $nextseq ${name}.fastq.gz + trim_galore --cores $cores --fastqc --gzip $c_r1 $tpc_r1 $nextseq ${name}.fastq.gz """ } else { """ [ ! -f ${name}_1.fastq.gz ] && ln -s ${reads[0]} ${name}_1.fastq.gz [ ! -f ${name}_2.fastq.gz ] && ln -s ${reads[1]} ${name}_2.fastq.gz - trim_galore --cores $task.cpus --paired --fastqc --gzip $c_r1 $c_r2 $tpc_r1 $tpc_r2 $nextseq ${name}_1.fastq.gz ${name}_2.fastq.gz + trim_galore --cores $cores --paired --fastqc --gzip $c_r1 $c_r2 $tpc_r1 $tpc_r2 $nextseq ${name}_1.fastq.gz ${name}_2.fastq.gz """ } } From 72fd092beca2fbe36cfed727080c1a41612bbec7 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Mon, 25 Nov 2019 17:58:44 +0000 Subject: [PATCH 037/226] Remove skip_plot_profile from test.config --- conf/test.config | 4 ---- 1 file changed, 4 deletions(-) diff --git a/conf/test.config b/conf/test.config index 9b712466..0435d184 100755 --- a/conf/test.config +++ b/conf/test.config @@ -28,8 +28,4 @@ params { // For speed to avoid CI time-out fingerprint_bins = 100 - - // MultiQC v1.7 bug with Python3 - // See https://github.com/ewels/MultiQC/issues/1057 - skip_plot_profile = true } From 520d3998ddabc2d81ad32f33264b3d0d79fbf59b Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 12 Dec 2019 11:51:20 +0000 Subject: [PATCH 038/226] Remove awsbatch.config --- conf/awsbatch.config | 18 ------------------ 1 file changed, 18 deletions(-) delete mode 100755 conf/awsbatch.config diff --git a/conf/awsbatch.config b/conf/awsbatch.config deleted file mode 100755 index 14af5866..00000000 --- a/conf/awsbatch.config +++ /dev/null @@ -1,18 +0,0 @@ -/* - * ------------------------------------------------- - * Nextflow config file for running on AWS batch - * ------------------------------------------------- - * Base config needed for running with -profile awsbatch - */ -params { - config_profile_name = 'AWSBATCH' - config_profile_description = 'AWSBATCH Cloud Profile' - config_profile_contact = 'Alexander Peltzer (@apeltzer)' - config_profile_url = 'https://aws.amazon.com/de/batch/' -} - -aws.region = params.awsregion -process.executor = 'awsbatch' -process.queue = params.awsqueue -executor.awscli = '/home/ec2-user/miniconda/bin/aws' -params.tracedir = './' From 572debc0f95c0cbf63fb63f9e72dca16dc600921 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 12 Dec 2019 14:16:27 +0000 Subject: [PATCH 039/226] Move awsbatch to configs --- docs/usage.md | 9 ++++++--- 1 file changed, 6 insertions(+), 3 deletions(-) diff --git a/docs/usage.md b/docs/usage.md index 704fa679..ebbe3520 100755 --- a/docs/usage.md +++ b/docs/usage.md @@ -51,6 +51,7 @@ * [AWS Batch specific parameters](#aws-batch-specific-parameters) * [`--awsqueue`](#--awsqueue) * [`--awsregion`](#--awsregion) + * [`--awscli`](#--awscli) * [Other command line parameters](#other-command-line-parameters) * [`--outdir`](#--outdir) * [`--email`](#--email) @@ -122,8 +123,6 @@ Use this parameter to choose a configuration profile. Profiles can give configur If `-profile` is not specified at all the pipeline will be run locally and expects all software to be installed and available on the `PATH`. -* `awsbatch` - * A generic configuration profile to be used with AWS Batch. * `conda` * A generic configuration profile to be used with [conda](https://conda.io/docs/) * Pulls most software from [Bioconda](https://bioconda.github.io/) @@ -424,7 +423,7 @@ If you have any questions or issues please send us a message on [Slack](https:// ## AWS Batch specific parameters -Running the pipeline on AWS Batch requires a couple of specific parameters to be set according to your AWS Batch configuration. Please use the `-awsbatch` profile and then specify all of the following parameters. +Running the pipeline on AWS Batch requires a couple of specific parameters to be set according to your AWS Batch configuration. Please use [`-profile awsbatch`](https://github.com/nf-core/configs/blob/master/conf/awsbatch.config) and then specify all of the following parameters. ### `--awsqueue` @@ -434,6 +433,10 @@ The JobQueue that you intend to use on AWS Batch. The AWS region to run your job in. Default is set to `eu-west-1` but can be adjusted to your needs. +### `--awscli` + +The [AWS CLI](https://www.nextflow.io/docs/latest/awscloud.html#aws-cli-installation) path in your custom AMI. Default: `/home/ec2-user/miniconda/bin/aws`. + Please make sure to also set the `-w/--work-dir` and `--outdir` parameters to a S3 storage bucket of your choice - you'll get an error message notifying you if you didn't. ## Other command line parameters From a8ebb1d074d830ff3fb3c59e459e041fc6dc9d55 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 12 Dec 2019 14:16:31 +0000 Subject: [PATCH 040/226] Move awsbatch to configs --- main.nf | 8 +++++--- 1 file changed, 5 insertions(+), 3 deletions(-) diff --git a/main.nf b/main.nf index 45c0e2bc..1125d257 100644 --- a/main.nf +++ b/main.nf @@ -84,6 +84,7 @@ def helpMessage() { AWSBatch --awsqueue [str] The AWSBatch JobQueue that needs to be set when running on AWSBatch --awsregion [str] The AWS Region for your AWS Batch job to run on + --awscli [str] Path to the AWS CLI tool """.stripIndent() } @@ -265,9 +266,10 @@ summary['Launch Dir'] = workflow.launchDir summary['Working Dir'] = workflow.workDir summary['Script Dir'] = workflow.projectDir summary['User'] = workflow.userName -if (workflow.profile == 'awsbatch') { +if (workflow.profile.contains('awsbatch')) { summary['AWS Region'] = params.awsregion summary['AWS Queue'] = params.awsqueue + summary['AWS CLI'] = params.awscli } summary['Config Profile'] = workflow.profile if (params.config_profile_description) summary['Config Description'] = params.config_profile_description @@ -554,7 +556,7 @@ if (params.skip_trimming) { cores = tcores } } - + // Added soft-links to original fastqs for consistent naming in MultiQC c_r1 = params.clip_r1 > 0 ? "--clip_r1 ${params.clip_r1}" : '' @@ -562,7 +564,7 @@ if (params.skip_trimming) { tpc_r1 = params.three_prime_clip_r1 > 0 ? "--three_prime_clip_r1 ${params.three_prime_clip_r1}" : '' tpc_r2 = params.three_prime_clip_r2 > 0 ? "--three_prime_clip_r2 ${params.three_prime_clip_r2}" : '' nextseq = params.trim_nextseq > 0 ? "--nextseq ${params.trim_nextseq}" : '' - + // Added soft-links to original fastqs for consistent naming in MultiQC if (params.single_end) { """ From c896ff68e630057f1ea306e560d0f6fb4c1694a4 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 12 Dec 2019 14:16:35 +0000 Subject: [PATCH 041/226] Move awsbatch to configs --- nextflow.config | 5 ----- 1 file changed, 5 deletions(-) diff --git a/nextflow.config b/nextflow.config index ceface2e..76db551d 100755 --- a/nextflow.config +++ b/nextflow.config @@ -52,10 +52,6 @@ params { skip_igv = false skip_multiqc = false - // Options: AWSBatch - awsqueue = false - awsregion = 'eu-west-1' - // Options: Config multiqc_config = "$baseDir/assets/multiqc/multiqc_config.yaml" bamtools_filter_pe_config = "$baseDir/assets/bamtools_filter_pe.json" @@ -105,7 +101,6 @@ try { } profiles { - awsbatch { includeConfig 'conf/awsbatch.config' } conda { process.conda = "$baseDir/environment.yml" } debug { process.beforeScript = 'echo $HOSTNAME' } docker { docker.enabled = true } From c3ac053abea214bf50ddaf675a9620d9077b816d Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 12 Dec 2019 14:38:07 +0000 Subject: [PATCH 042/226] Update TrimGalore cores calculation --- main.nf | 11 +++++------ 1 file changed, 5 insertions(+), 6 deletions(-) diff --git a/main.nf b/main.nf index 1125d257..e219fa5d 100644 --- a/main.nf +++ b/main.nf @@ -548,16 +548,15 @@ if (params.skip_trimming) { script: // Calculate number of --cores for TrimGalore based on value of task.cpus // See: https://github.com/FelixKrueger/TrimGalore/blob/master/Changelog.md#version-060-release-on-1-mar-2019 - // See: https://github.com/nf-core/atacseq/pull/65#issuecomment-557839116 + // See: https://github.com/nf-core/atacseq/pull/65 def cores = 1 if (task.cpus) { - tcores = (((task.cpus as int) - 3) / 3) as int - if (tcores > 1) { - cores = tcores - } + cores = (task.cpus as int) - 4 + if (params.single_end) cores = (task.cpus as int) - 3 + if (cores < 1) cores = 1 + if (cores > 4) cores = 4 } - // Added soft-links to original fastqs for consistent naming in MultiQC c_r1 = params.clip_r1 > 0 ? "--clip_r1 ${params.clip_r1}" : '' c_r2 = params.clip_r2 > 0 ? "--clip_r2 ${params.clip_r2}" : '' From 10169489c96d7c961b5e249342a9a4a16e9b5508 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 13 Dec 2019 15:58:03 +0000 Subject: [PATCH 043/226] Update via template --- .github/CONTRIBUTING.md | 52 ++++++++++++++++++++++++----------------- 1 file changed, 31 insertions(+), 21 deletions(-) diff --git a/.github/CONTRIBUTING.md b/.github/CONTRIBUTING.md index 47dc0f76..0d1890fa 100644 --- a/.github/CONTRIBUTING.md +++ b/.github/CONTRIBUTING.md @@ -1,47 +1,57 @@ # nf-core/atacseq: Contributing Guidelines -Hi there! Many thanks for taking an interest in improving nf-core/atacseq. +Hi there! +Many thanks for taking an interest in improving nf-core/atacseq. -We try to manage the required tasks for nf-core/atacseq using GitHub issues, you probably came to this page when creating one. Please use the pre-filled template to save time. - -However, don't be put off by this template - other more general issues and suggestions are welcome! Contributions to the code are even more welcome ;) - -> If you need help using or modifying nf-core/atacseq then the best place to ask is on the pipeline channel on [Slack](https://nf-co.re/join/slack/). +We try to manage the required tasks for nf-core/atacseq using GitHub issues, you probably came to this page when creating one. +Please use the pre-filled template to save time. +However, don't be put off by this template - other more general issues and suggestions are welcome! +Contributions to the code are even more welcome ;) +> If you need help using or modifying nf-core/atacseq then the best place to ask is on the nf-core Slack [#atacseq](https://nfcore.slack.com/channels/atacseq) channel ([join our Slack here](https://nf-co.re/join/slack)). ## Contribution workflow -If you'd like to write some code for nf-core/atacseq, the standard workflow -is as follows: -1. Check that there isn't already an issue about your idea in the - [nf-core/atacseq issues](https://github.com/nf-core/atacseq/issues) to avoid - duplicating work. +If you'd like to write some code for nf-core/atacseq, the standard workflow is as follows: + +1. Check that there isn't already an issue about your idea in the [nf-core/atacseq issues](https://github.com/nf-core/atacseq/issues) to avoid duplicating work * If there isn't one already, please create one so that others know you're working on this -2. Fork the [nf-core/atacseq repository](https://github.com/nf-core/atacseq) to your GitHub account +2. [Fork](https://help.github.com/en/github/getting-started-with-github/fork-a-repo) the [nf-core/atacseq repository](https://github.com/nf-core/atacseq) to your GitHub account 3. Make the necessary changes / additions within your forked repository -4. Submit a Pull Request against the `dev` branch and wait for the code to be reviewed and merged. - -If you're not used to this workflow with git, you can start with some [basic docs from GitHub](https://help.github.com/articles/fork-a-repo/) or even their [excellent interactive tutorial](https://try.github.io/). +4. Submit a Pull Request against the `dev` branch and wait for the code to be reviewed and merged +If you're not used to this workflow with git, you can start with some [docs from GitHub](https://help.github.com/en/github/collaborating-with-issues-and-pull-requests) or even their [excellent `git` resources](https://try.github.io/). ## Tests -When you create a pull request with changes, [Travis CI](https://travis-ci.com/) will run automatic tests. + +When you create a pull request with changes, [GitHub Actions](https://github.com/features/actions) will run automatic tests. Typically, pull-requests are only fully reviewed when these tests are passing, though of course we can help out before then. There are typically two types of tests that run: ### Lint Tests -The nf-core has a [set of guidelines](https://nf-co.re/developers/guidelines) which all pipelines must adhere to. + +`nf-core` has a [set of guidelines](https://nf-co.re/developers/guidelines) which all pipelines must adhere to. To enforce these and ensure that all pipelines stay in sync, we have developed a helper tool which runs checks on the pipeline code. This is in the [nf-core/tools repository](https://github.com/nf-core/tools) and once installed can be run locally with the `nf-core lint ` command. If any failures or warnings are encountered, please follow the listed URL for more documentation. ### Pipeline Tests -Each nf-core pipeline should be set up with a minimal set of test-data. -Travis CI then runs the pipeline on this data to ensure that it exists successfully. + +Each `nf-core` pipeline should be set up with a minimal set of test-data. +`GitHub Actions` then runs the pipeline on this data to ensure that it exits successfully. If there are any failures then the automated tests fail. -These tests are run both with the latest available version of Nextflow and also the minimum required version that is stated in the pipeline code. +These tests are run both with the latest available version of `Nextflow` and also the minimum required version that is stated in the pipeline code. + +## Patch + +: warning: Only in the unlikely and regretful event of a release happening with a bug. + +* On your own fork, make a new branch `patch` based on `upstream/master`. +* Fix the bug, and bump version (X.Y.Z+1). +* A PR should be made on `master` from patch to directly this particular bug. ## Getting help -For further information/help, please consult the [nf-core/atacseq documentation](https://github.com/nf-core/atacseq#documentation) and don't hesitate to get in touch on the [nf-core/atacseq pipeline channel](https://nfcore.slack.com/channels/atacseq) on [Slack](https://nf-co.re/join/slack/). + +For further information/help, please consult the [nf-core/atacseq documentation](https://nf-co.re/nf-core/atacseq/docs) and don't hesitate to get in touch on the nf-core Slack [#atacseq](https://nfcore.slack.com/channels/atacseq) channel ([join our Slack here](https://nf-co.re/join/slack)). From b7ef415fee1164d734d910756eae5e7180f65bba Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 13 Dec 2019 15:58:17 +0000 Subject: [PATCH 044/226] Update via template --- .github/ISSUE_TEMPLATE/bug_report.md | 43 +++++++++++++++++----------- 1 file changed, 27 insertions(+), 16 deletions(-) diff --git a/.github/ISSUE_TEMPLATE/bug_report.md b/.github/ISSUE_TEMPLATE/bug_report.md index b287bce7..010263cf 100644 --- a/.github/ISSUE_TEMPLATE/bug_report.md +++ b/.github/ISSUE_TEMPLATE/bug_report.md @@ -1,31 +1,42 @@ +# nf-core/atacseq bug report + Hi there! -Thanks for telling us about a problem with the pipeline. Please delete this text and anything that's not relevant from the template below: +Thanks for telling us about a problem with the pipeline. +Please delete this text and anything that's not relevant from the template below: + +## Describe the bug -#### Describe the bug A clear and concise description of what the bug is. -#### Steps to reproduce +## Steps to reproduce + Steps to reproduce the behaviour: + 1. Command line: `nextflow run ...` 2. See error: _Please provide your error message_ -#### Expected behaviour +## Expected behaviour + A clear and concise description of what you expected to happen. -#### System: - - Hardware: [e.g. HPC, Desktop, Cloud...] - - Executor: [e.g. slurm, local, awsbatch...] - - OS: [e.g. CentOS Linux, macOS, Linux Mint...] - - Version [e.g. 7, 10.13.6, 18.3...] +## System + +- Hardware: +- Executor: +- OS: +- Version + +## Nextflow Installation + +- Version: + +## Container engine -#### Nextflow Installation: - - Version: [e.g. 0.31.0] +- Engine: +- version: +- Image tag: -#### Container engine: - - Engine: [e.g. Conda, Docker or Singularity] - - version: [e.g. 1.0.0] - - Image tag: [e.g. nfcore/atacseq:1.0.0] +## Additional context -#### Additional context Add any other context about the problem here. From 4e5538a9298e49b08d7b41912975f3535e1b601c Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 13 Dec 2019 15:58:31 +0000 Subject: [PATCH 045/226] Update via template --- .github/ISSUE_TEMPLATE/feature_request.md | 18 +++++++++++++----- 1 file changed, 13 insertions(+), 5 deletions(-) diff --git a/.github/ISSUE_TEMPLATE/feature_request.md b/.github/ISSUE_TEMPLATE/feature_request.md index 1f025b77..222fe51a 100644 --- a/.github/ISSUE_TEMPLATE/feature_request.md +++ b/.github/ISSUE_TEMPLATE/feature_request.md @@ -1,16 +1,24 @@ +# nf-core/atacseq feature request + Hi there! -Thanks for suggesting a new feature for the pipeline! Please delete this text and anything that's not relevant from the template below: +Thanks for suggesting a new feature for the pipeline! +Please delete this text and anything that's not relevant from the template below: + +## Is your feature request related to a problem? Please describe -#### Is your feature request related to a problem? Please describe. A clear and concise description of what the problem is. + Ex. I'm always frustrated when [...] -#### Describe the solution you'd like +## Describe the solution you'd like + A clear and concise description of what you want to happen. -#### Describe alternatives you've considered +## Describe alternatives you've considered + A clear and concise description of any alternative solutions or features you've considered. -#### Additional context +## Additional context + Add any other context about the feature request here. From 8decb44d324c10677999ba285af9d811844aec61 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 13 Dec 2019 15:58:46 +0000 Subject: [PATCH 046/226] Update via template --- .github/PULL_REQUEST_TEMPLATE.md | 26 +++++++++++++++----------- 1 file changed, 15 insertions(+), 11 deletions(-) diff --git a/.github/PULL_REQUEST_TEMPLATE.md b/.github/PULL_REQUEST_TEMPLATE.md index ab7fd57d..67681d0c 100644 --- a/.github/PULL_REQUEST_TEMPLATE.md +++ b/.github/PULL_REQUEST_TEMPLATE.md @@ -1,15 +1,19 @@ +# nf-core/atacseq pull request + Many thanks for contributing to nf-core/atacseq! -Please fill in the appropriate checklist below (delete whatever is not relevant). These are the most common things requested on pull requests (PRs). +Please fill in the appropriate checklist below (delete whatever is not relevant). +These are the most common things requested on pull requests (PRs). ## PR checklist - - [ ] This comment contains a description of changes (with reason) - - [ ] If you've fixed a bug or added code that should be tested, add tests! - - [ ] If necessary, also make a PR on the [nf-core/atacseq branch on the nf-core/test-datasets repo]( https://github.com/nf-core/test-datasets/pull/new/nf-core/atacseq) - - [ ] Ensure the test suite passes (`nextflow run . -profile test,docker`). - - [ ] Make sure your code lints (`nf-core lint .`). - - [ ] Documentation in `docs` is updated - - [ ] `CHANGELOG.md` is updated - - [ ] `README.md` is updated - -**Learn more about contributing:** https://github.com/nf-core/atacseq/tree/master/.github/CONTRIBUTING.md + +- [ ] This comment contains a description of changes (with reason) +- [ ] If you've fixed a bug or added code that should be tested, add tests! +- [ ] If necessary, also make a PR on the [nf-core/atacseq branch on the nf-core/test-datasets repo](https://github.com/nf-core/test-datasets/pull/new/nf-core/atacseq) +- [ ] Ensure the test suite passes (`nextflow run . -profile test,docker`). +- [ ] Make sure your code lints (`nf-core lint .`). +- [ ] Documentation in `docs` is updated +- [ ] `CHANGELOG.md` is updated +- [ ] `README.md` is updated + +**Learn more about contributing:** [CONTRIBUTING.md](https://github.com/nf-core/atacseq/tree/master/.github/CONTRIBUTING.md) \ No newline at end of file From 9674450ed891f03dde5d4bbb36f13793df842f50 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 13 Dec 2019 15:59:03 +0000 Subject: [PATCH 047/226] Update test dir --- .gitignore | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/.gitignore b/.gitignore index 5b54e3e6..0189a444 100755 --- a/.gitignore +++ b/.gitignore @@ -3,5 +3,5 @@ work/ data/ results/ .DS_Store -tests/test_data +test* *.pyc From d8b2790f5becace0aa92be183d26a04fea112750 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 13 Dec 2019 15:59:16 +0000 Subject: [PATCH 048/226] Add NF tower --- .travis.yml | 13 +++++++++---- 1 file changed, 9 insertions(+), 4 deletions(-) diff --git a/.travis.yml b/.travis.yml index 55073b54..d3c40257 100755 --- a/.travis.yml +++ b/.travis.yml @@ -9,7 +9,7 @@ matrix: before_install: # PRs to master are only ok if coming from dev branch - - '[ $TRAVIS_PULL_REQUEST = "false" ] || [ $TRAVIS_BRANCH != "master" ] || ([ $TRAVIS_PULL_REQUEST_SLUG = $TRAVIS_REPO_SLUG ] && ([ $TRAVIS_PULL_REQUEST_BRANCH = "dev" ] || [ $TRAVIS_PULL_REQUEST_BRANCH = "patch" ]))' + - '[ $TRAVIS_PULL_REQUEST = "false" ] || [ $TRAVIS_BRANCH != "master" ] || ([ $TRAVIS_PULL_REQUEST_SLUG = $TRAVIS_REPO_SLUG ] && [ $TRAVIS_PULL_REQUEST_BRANCH = "dev" ]) || [ $TRAVIS_PULL_REQUEST_BRANCH = "patch" ]' # Pull the docker image first so the test doesn't wait for this - docker pull nfcore/atacseq:dev # Fake the tag locally so that the pipeline runs properly @@ -30,8 +30,13 @@ install: - sudo apt-get install npm && npm install -g markdownlint-cli env: - - NXF_VER='19.10.0' # Specify a minimum NF version that should be tested and work - - NXF_VER='' # Plus: get the latest NF version and check that it works + # Tower token is to inspect runs on https://tower.nf + # Use public mailbox nf-core@mailinator.com to log in: https://www.mailinator.com/v3/index.jsp?zone=public&query=nf-core + # Specify a minimum NF version that should be tested and work + - NXF_VER='19.10.0' TOWER_ACCESS_TOKEN="1c1f493bc2703472d6f1b9f6fb9e9d117abab7b1" + # Plus: get the latest NF version and check that it works + - NXF_VER='' TOWER_ACCESS_TOKEN="1c1f493bc2703472d6f1b9f6fb9e9d117abab7b1" + script: # Lint the pipeline code @@ -39,4 +44,4 @@ script: # Lint the documentation - markdownlint ${TRAVIS_BUILD_DIR} -c ${TRAVIS_BUILD_DIR}/.github/markdownlint.yml # Run the pipeline with the test profile - - nextflow run ${TRAVIS_BUILD_DIR} -profile test,docker -ansi-log false + - nextflow run ${TRAVIS_BUILD_DIR} -profile test,docker -ansi-log false -name atacseq-${TRAVIS_EVENT_TYPE}-${TRAVIS_PULL_REQUEST}-${TRAVIS_COMMIT:0:6}-test-description From 86c8f55014209d7ad632f83675cbcc8668d79844 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 13 Dec 2019 15:59:30 +0000 Subject: [PATCH 049/226] Update docker options --- nextflow.config | 20 ++++++++++++-------- 1 file changed, 12 insertions(+), 8 deletions(-) diff --git a/nextflow.config b/nextflow.config index 76db551d..8941e96d 100755 --- a/nextflow.config +++ b/nextflow.config @@ -103,17 +103,21 @@ try { profiles { conda { process.conda = "$baseDir/environment.yml" } debug { process.beforeScript = 'echo $HOSTNAME' } - docker { docker.enabled = true } - singularity { singularity.enabled = true - singularity.autoMounts = true } + docker { + docker.enabled = true + // Avoid this error: + // WARNING: Your kernel does not support swap limit capabilities or the cgroup is not mounted. Memory limited without swap. + // Testing this in nf-core after discussion here https://github.com/nf-core/tools/pull/351 + // once this is established and works well, nextflow might implement this behavior as new default. + docker.runOptions = '-u \$(id -u):\$(id -g)' + } + singularity { + singularity.enabled = true + singularity.autoMounts = true + } test { includeConfig 'conf/test.config' } } -// Avoid this error: -// WARNING: Your kernel does not support swap limit capabilities or the cgroup is not mounted. Memory limited without swap. -// Testing this in nf-core after discussion here https://github.com/nf-core/tools/pull/351, once this is established and works well, nextflow might implement this behavior as new default. -docker.runOptions = '-u \$(id -u):\$(id -g)' - // Load igenomes.config if required if (!params.igenomes_ignore) { includeConfig 'conf/igenomes.config' From ddad9de909e5086ed7b8f4fe7b9db4251aefdd93 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 13 Dec 2019 15:59:48 +0000 Subject: [PATCH 050/226] Change name --- assets/email_template.html | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/assets/email_template.html b/assets/email_template.html index 61a98cb6..098cd4bd 100755 --- a/assets/email_template.html +++ b/assets/email_template.html @@ -5,7 +5,7 @@ - + nf-core/atacseq Pipeline Report From ff930e7a3c3dc27a93ec49d8b5bd124966993400 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 13 Dec 2019 16:00:03 +0000 Subject: [PATCH 051/226] Adjust spacing --- assets/email_template.txt | 1 + 1 file changed, 1 insertion(+) diff --git a/assets/email_template.txt b/assets/email_template.txt index 5f89db73..40533389 100755 --- a/assets/email_template.txt +++ b/assets/email_template.txt @@ -28,6 +28,7 @@ The command used to launch the workflow was as follows: $commandLine + Pipeline Configuration: ----------------------- <% out << summary.collect{ k,v -> " - $k: $v" }.join("\n") %> From cd80d0ea9dbd6f15386260a40d0ab04600c23674 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 13 Dec 2019 16:00:14 +0000 Subject: [PATCH 052/226] Fix slack link --- CODE_OF_CONDUCT.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/CODE_OF_CONDUCT.md b/CODE_OF_CONDUCT.md index 1cda7600..cf930c8a 100755 --- a/CODE_OF_CONDUCT.md +++ b/CODE_OF_CONDUCT.md @@ -34,7 +34,7 @@ This Code of Conduct applies both within project spaces and in public spaces whe ## Enforcement -Instances of abusive, harassing, or otherwise unacceptable behavior may be reported by contacting the project team on [Slack](https://nf-co.re/join/slack/). The project team will review and investigate all complaints, and will respond in a way that it deems appropriate to the circumstances. The project team is obligated to maintain confidentiality with regard to the reporter of an incident. Further details of specific enforcement policies may be posted separately. +Instances of abusive, harassing, or otherwise unacceptable behavior may be reported by contacting the project team on [Slack](https://nf-co.re/join/slack). The project team will review and investigate all complaints, and will respond in a way that it deems appropriate to the circumstances. The project team is obligated to maintain confidentiality with regard to the reporter of an incident. Further details of specific enforcement policies may be posted separately. Project maintainers who do not follow or enforce the Code of Conduct in good faith may face temporary or permanent repercussions as determined by other members of the project's leadership. From 0b739e291b4f223b6992cea32dca18b89089ed30 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 13 Dec 2019 16:00:23 +0000 Subject: [PATCH 053/226] Update description --- Dockerfile | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/Dockerfile b/Dockerfile index eed4f3cb..e9f6bfde 100755 --- a/Dockerfile +++ b/Dockerfile @@ -1,6 +1,6 @@ -FROM nfcore/base:1.7 +FROM nfcore/base:dev LABEL authors="Harshil Patel" \ - description="Docker image containing all requirements for nf-core/atacseq pipeline" + description="Docker image containing all software requirements for the nf-core/atacseq pipeline" # Install the conda environment COPY environment.yml / From 7b822c82809a0bdb7ec76463c12f969cf8cd2f27 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 13 Dec 2019 16:00:33 +0000 Subject: [PATCH 054/226] Update badges --- README.md | 2 ++ 1 file changed, 2 insertions(+) diff --git a/README.md b/README.md index 32f5bb93..eab86f7f 100644 --- a/README.md +++ b/README.md @@ -1,6 +1,8 @@ # ![nfcore/atacseq](docs/images/nf-core-atacseq_logo.png) [![Build Status](https://travis-ci.com/nf-core/atacseq.svg?branch=master)](https://travis-ci.com/nf-core/atacseq) +[![GitHub Actions CI Status](https://github.com/nf-core/atacseq/workflows/nf-core%20CI/badge.svg)](https://github.com/nf-core/atacseq/actions) +[![GitHub Actions Linting Status](https://github.com/nf-core/atacseq/workflows/nf-core%20linting/badge.svg)](https://github.com/nf-core/atacseq/actions) [![Nextflow](https://img.shields.io/badge/nextflow-%E2%89%A519.10.0-brightgreen.svg)](https://www.nextflow.io/) [![install with bioconda](https://img.shields.io/badge/install%20with-bioconda-brightgreen.svg)](http://bioconda.github.io/) From 0e0b38c35c8d636c1b4753f9e27c6ebc10cf0438 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 13 Dec 2019 16:00:51 +0000 Subject: [PATCH 055/226] Add GitHub actions --- .github/workflows/branch.yml | 16 ++++++++++++++ .github/workflows/ci.yml | 22 +++++++++++++++++++ .github/workflows/linting.yml | 41 +++++++++++++++++++++++++++++++++++ 3 files changed, 79 insertions(+) create mode 100755 .github/workflows/branch.yml create mode 100755 .github/workflows/ci.yml create mode 100755 .github/workflows/linting.yml diff --git a/.github/workflows/branch.yml b/.github/workflows/branch.yml new file mode 100755 index 00000000..0df3a69a --- /dev/null +++ b/.github/workflows/branch.yml @@ -0,0 +1,16 @@ +name: nf-core branch protection +# This workflow is triggered on PRs to master branch on the repository +# It fails when someone tries to make a PR against the nf-core `master` branch instead of `dev` +on: + pull_request: + branches: + - master + +jobs: + test: + runs-on: ubuntu-18.04 + steps: + # PRs are only ok if coming from an nf-core `dev` branch or a fork `patch` branch + - name: Check PRs + run: | + { [[ $(git remote get-url origin) == *nf-core-atacseq ]] && [[ ${GITHUB_HEAD_REF} = "dev" ]]; } || [[ ${GITHUB_HEAD_REF} == "patch" ]] diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml new file mode 100755 index 00000000..7a692d18 --- /dev/null +++ b/.github/workflows/ci.yml @@ -0,0 +1,22 @@ +name: nf-core CI +# This workflow is triggered on pushes and PRs to the repository. +# It runs the pipeline with the minimal test dataset to check that it completes without any syntax errors +on: [push, pull_request] + +jobs: + test: + runs-on: ubuntu-18.04 + strategy: + matrix: + # Nextflow versions: check pipeline minimum and current latest + nxf_ver: ['19.10.0', ''] + steps: + - uses: actions/checkout@v1 + - name: Install Nextflow + run: | + export NXF_VER=${{ matrix.nxf_ver }} + wget -qO- get.nextflow.io | bash + sudo mv nextflow /usr/local/bin/ + - name: Run test + run: | + nextflow run ${GITHUB_WORKSPACE} -profile test,docker diff --git a/.github/workflows/linting.yml b/.github/workflows/linting.yml new file mode 100755 index 00000000..7354dc74 --- /dev/null +++ b/.github/workflows/linting.yml @@ -0,0 +1,41 @@ +name: nf-core linting +# This workflow is triggered on pushes and PRs to the repository. +# It runs the `nf-core lint` and markdown lint tests to ensure that the code meets the nf-core guidelines +on: [push, pull_request] + +jobs: + Markdown: + runs-on: ubuntu-18.04 + steps: + - uses: actions/checkout@v1 + - uses: actions/setup-node@v1 + with: + node-version: '10' + - name: Install markdownlint + run: | + npm install -g markdownlint-cli + - name: Run Markdownlint + run: | + markdownlint ${GITHUB_WORKSPACE} -c ${GITHUB_WORKSPACE}/.github/markdownlint.yml + nf-core: + runs-on: ubuntu-latest + steps: + - uses: actions/checkout@v1 + - name: Install Nextflow + run: | + wget -qO- get.nextflow.io | bash + sudo mv nextflow /usr/local/bin/ + - uses: actions/setup-python@v1 + with: + python-version: '3.6' + architecture: 'x64' + - name: Install pip + run: | + sudo apt install python3-pip + pip install --upgrade pip + - name: Install nf-core tools + run: | + pip install nf-core + - name: Run nf-core lint + run: | + nf-core lint ${GITHUB_WORKSPACE} From a5756abced0a304497d46ed6909a4245bcf7560d Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 13 Dec 2019 17:37:19 +0000 Subject: [PATCH 056/226] Disable ANSI --- .github/workflows/ci.yml | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml index 7a692d18..75fc7281 100755 --- a/.github/workflows/ci.yml +++ b/.github/workflows/ci.yml @@ -1,7 +1,7 @@ name: nf-core CI # This workflow is triggered on pushes and PRs to the repository. # It runs the pipeline with the minimal test dataset to check that it completes without any syntax errors -on: [push, pull_request] +on: [push, pull_request] jobs: test: @@ -19,4 +19,4 @@ jobs: sudo mv nextflow /usr/local/bin/ - name: Run test run: | - nextflow run ${GITHUB_WORKSPACE} -profile test,docker + nextflow run ${GITHUB_WORKSPACE} -ansi-log false -profile test,docker From 8e59b701a614d43cacc2d6ace108a2cada682219 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sat, 14 Dec 2019 10:53:43 +0000 Subject: [PATCH 057/226] Revert branch test --- .travis.yml | 3 ++- 1 file changed, 2 insertions(+), 1 deletion(-) diff --git a/.travis.yml b/.travis.yml index d3c40257..3b07ff58 100755 --- a/.travis.yml +++ b/.travis.yml @@ -9,7 +9,8 @@ matrix: before_install: # PRs to master are only ok if coming from dev branch - - '[ $TRAVIS_PULL_REQUEST = "false" ] || [ $TRAVIS_BRANCH != "master" ] || ([ $TRAVIS_PULL_REQUEST_SLUG = $TRAVIS_REPO_SLUG ] && [ $TRAVIS_PULL_REQUEST_BRANCH = "dev" ]) || [ $TRAVIS_PULL_REQUEST_BRANCH = "patch" ]' + #- '[ $TRAVIS_PULL_REQUEST = "false" ] || [ $TRAVIS_BRANCH != "master" ] || ([ $TRAVIS_PULL_REQUEST_SLUG = $TRAVIS_REPO_SLUG ] && [ $TRAVIS_PULL_REQUEST_BRANCH = "dev" ]) || [ $TRAVIS_PULL_REQUEST_BRANCH = "patch" ]' + - '[ $TRAVIS_PULL_REQUEST = "false" ] || [ $TRAVIS_BRANCH != "master" ] || ([ $TRAVIS_PULL_REQUEST_SLUG = $TRAVIS_REPO_SLUG ] && ([ $TRAVIS_PULL_REQUEST_BRANCH = "dev" ] || [ $TRAVIS_PULL_REQUEST_BRANCH = "patch" ]))' # Pull the docker image first so the test doesn't wait for this - docker pull nfcore/atacseq:dev # Fake the tag locally so that the pipeline runs properly From fc7cc29664570777c58badc9e02eeb32f05cbcb6 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sat, 14 Dec 2019 10:54:01 +0000 Subject: [PATCH 058/226] Revert Dockerfile string --- Dockerfile | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/Dockerfile b/Dockerfile index e9f6bfde..abb90542 100755 --- a/Dockerfile +++ b/Dockerfile @@ -1,4 +1,4 @@ -FROM nfcore/base:dev +FROM nfcore/base:1.7 LABEL authors="Harshil Patel" \ description="Docker image containing all software requirements for the nf-core/atacseq pipeline" From c383ff6a7566ce0e409b051f1a91f2414ac342fc Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sat, 14 Dec 2019 22:54:02 +0000 Subject: [PATCH 059/226] Update CHANGELOG --- CHANGELOG.md | 5 +++++ 1 file changed, 5 insertions(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index fc18f302..b2e48240 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -10,6 +10,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Added` * [#63](https://github.com/nf-core/atacseq/issues/63) - Added multicore support for Trim Galore! +* Added GitHub Actions CI tests ### `Fixed` @@ -44,6 +45,10 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Deprecated` +| Deprecated | Replacement | +|------------------------------|---------------------------| +| `--skip_diff_analysis` | `--skip_consensus_peaks` | + ## [1.1.0] - 2019-11-05 ### `Added` From 7580248f709b724c702e9933b58d06d765cf5478 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sat, 14 Dec 2019 22:54:09 +0000 Subject: [PATCH 060/226] Replace --skip_diff_analysis with --skip_consensus_peaks --- docs/usage.md | 6 +++--- 1 file changed, 3 insertions(+), 3 deletions(-) diff --git a/docs/usage.md b/docs/usage.md index ebbe3520..3e51475a 100755 --- a/docs/usage.md +++ b/docs/usage.md @@ -43,7 +43,7 @@ * [`--broad_cutoff`](#--broad_cutoff) * [`--min_reps_consensus`](#--min_reps_consensus) * [`--save_macs_pileup`](#--save_macs_pileup) - * [`--skip_diff_analysis`](#--skip_diff_analysis) + * [`--skip_consensus_peaks`](#--skip_consensus_peaks) * [Skipping QC steps](#skipping-qc-steps) * [Job resources](#job-resources) * [Automatic resubmission](#automatic-resubmission) @@ -387,9 +387,9 @@ Number of biological replicates required from a given condition for a peak to co Instruct MACS2 to create bedGraph files using the `-B --SPMR` parameters. -### `--skip_diff_analysis` +### `--skip_consensus_peaks` -Skip read counting and differential analysis step. +Skip consensus peak generation and differential binding analysis. ## Skipping QC steps From 79a0b7ad57f84f12edb8809affb49e94b308f5bf Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sat, 14 Dec 2019 22:54:13 +0000 Subject: [PATCH 061/226] Replace --skip_diff_analysis with --skip_consensus_peaks --- main.nf | 16 ++++++++-------- 1 file changed, 8 insertions(+), 8 deletions(-) diff --git a/main.nf b/main.nf index e219fa5d..bb05a290 100644 --- a/main.nf +++ b/main.nf @@ -62,7 +62,7 @@ def helpMessage() { --broad_cutoff [float] Specifies broad cutoff value for MACS2. Only used when --narrow_peak isnt specified (Default: 0.1) --min_reps_consensus [int] Number of biological replicates required from a given condition for a peak to contribute to a consensus peak (Default: 1) --save_macs_pileup [bool] Instruct MACS2 to create bedGraph files normalised to signal per million reads - --skip_diff_analysis [bool] Skip differential binding analysis + --skip_consensus_peaks [bool] Skip consensus peak generation and differential binding analysis QC --skip_fastqc [bool] Skip FastQC @@ -250,7 +250,7 @@ if (params.save_trimmed) summary['Save Trimmed'] = 'Yes' if (params.save_align_intermeds) summary['Save Intermeds'] = 'Yes' if (params.save_macs_pileup) summary['Save MACS2 Pileup'] = 'Yes' if (params.skip_merge_replicates) summary['Skip Merge Replicates'] = 'Yes' -if (params.skip_diff_analysis) summary['Skip Diff Analysis'] = 'Yes' +if (params.skip_consensus_peaks) summary['Skip Consensus Peaks'] = 'Yes' if (params.skip_fastqc) summary['Skip FastQC'] = 'Yes' if (params.skip_picard_metrics) summary['Skip Picard Metrics'] = 'Yes' if (params.skip_preseq) summary['Skip Preseq'] = 'Yes' @@ -1194,7 +1194,7 @@ process MergedLibConsensusPeakSet { } when: - params.macs_gsize && (replicatesExist || multipleGroups) + params.macs_gsize && (replicatesExist || multipleGroups) && !params.skip_consensus_peaks input: file peaks from ch_mlib_macs_consensus.collect{ it[1] } @@ -1243,7 +1243,7 @@ process MergedLibConsensusPeakSetAnnotate { publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: 'copy' when: - params.macs_gsize && (replicatesExist || multipleGroups) + params.macs_gsize && (replicatesExist || multipleGroups) && !params.skip_consensus_peaks input: file bed from ch_mlib_macs_consensus_bed @@ -1282,7 +1282,7 @@ process MergedLibConsensusPeakSetDESeq { } when: - params.macs_gsize && replicatesExist && multipleGroups && !params.skip_diff_analysis + params.macs_gsize && replicatesExist && multipleGroups && !params.skip_consensus_peaks input: file bams from ch_mlib_name_bam_mlib_counts.collect{ it[1] } @@ -1661,7 +1661,7 @@ process MergedRepConsensusPeakSet { } when: - !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups + !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks input: file peaks from ch_mrep_macs_consensus.collect{ it[1] } @@ -1709,7 +1709,7 @@ process MergedRepConsensusPeakSetAnnotate { publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: 'copy' when: - !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups + !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks input: file bed from ch_mrep_macs_consensus_bed @@ -1748,7 +1748,7 @@ process MergedRepConsensusPeakSetDESeq { } when: - !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_diff_analysis + !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks input: file bams from ch_mlib_name_bam_mrep_counts.collect{ it[1] } From 4d4876bf55efeaf4037ce21092ab8f95ee786cd2 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sat, 14 Dec 2019 22:54:16 +0000 Subject: [PATCH 062/226] Replace --skip_diff_analysis with --skip_consensus_peaks --- nextflow.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/nextflow.config b/nextflow.config index 8941e96d..20733b75 100755 --- a/nextflow.config +++ b/nextflow.config @@ -40,7 +40,7 @@ params { broad_cutoff = 0.1 min_reps_consensus = 1 save_macs_pileup = false - skip_diff_analysis = false + skip_consensus_peaks = false // Options: QC skip_fastqc = false From 1adb3f3f46cabcfe754219db963e73b7aac23e7d Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sun, 15 Dec 2019 17:37:07 +0000 Subject: [PATCH 063/226] Fix branch name --- .github/workflows/branch.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/.github/workflows/branch.yml b/.github/workflows/branch.yml index 0df3a69a..9ba92076 100755 --- a/.github/workflows/branch.yml +++ b/.github/workflows/branch.yml @@ -13,4 +13,4 @@ jobs: # PRs are only ok if coming from an nf-core `dev` branch or a fork `patch` branch - name: Check PRs run: | - { [[ $(git remote get-url origin) == *nf-core-atacseq ]] && [[ ${GITHUB_HEAD_REF} = "dev" ]]; } || [[ ${GITHUB_HEAD_REF} == "patch" ]] + { [[ $(git remote get-url origin) == *nf-core/atacseq ]] && [[ ${GITHUB_HEAD_REF} = "dev" ]]; } || [[ ${GITHUB_HEAD_REF} == "patch" ]] From 0b0713952b9fb84ea3c68f1f1c5401f85cd8a5f6 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sun, 15 Dec 2019 17:44:32 +0000 Subject: [PATCH 064/226] Pull Docker image for CI tests --- .github/workflows/ci.yml | 5 ++++- 1 file changed, 4 insertions(+), 1 deletion(-) diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml index 75fc7281..913e8e10 100755 --- a/.github/workflows/ci.yml +++ b/.github/workflows/ci.yml @@ -14,9 +14,12 @@ jobs: - uses: actions/checkout@v1 - name: Install Nextflow run: | - export NXF_VER=${{ matrix.nxf_ver }} + {% raw %}export NXF_VER=${{ matrix.nxf_ver }}{% endraw %} wget -qO- get.nextflow.io | bash sudo mv nextflow /usr/local/bin/ + - name: Pull image + run: | + docker pull nfcore/atacseq:dev && docker tag nfcore/atacseq:dev nfcore/atacseq:dev - name: Run test run: | nextflow run ${GITHUB_WORKSPACE} -ansi-log false -profile test,docker From 179912c74be9c95ec9dd0360b32046b3ea1dc105 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sun, 15 Dec 2019 17:55:54 +0000 Subject: [PATCH 065/226] Remove cookiecutter tag --- .github/workflows/ci.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml index 913e8e10..39fc6c84 100755 --- a/.github/workflows/ci.yml +++ b/.github/workflows/ci.yml @@ -14,7 +14,7 @@ jobs: - uses: actions/checkout@v1 - name: Install Nextflow run: | - {% raw %}export NXF_VER=${{ matrix.nxf_ver }}{% endraw %} + export NXF_VER=${{ matrix.nxf_ver }} wget -qO- get.nextflow.io | bash sudo mv nextflow /usr/local/bin/ - name: Pull image From 786df1c3df34125321b5492c8aec7b1754d1bfa2 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Mon, 16 Dec 2019 17:59:42 +0000 Subject: [PATCH 066/226] Update origin glob --- .github/workflows/branch.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/.github/workflows/branch.yml b/.github/workflows/branch.yml index 9ba92076..37a3aa74 100755 --- a/.github/workflows/branch.yml +++ b/.github/workflows/branch.yml @@ -13,4 +13,4 @@ jobs: # PRs are only ok if coming from an nf-core `dev` branch or a fork `patch` branch - name: Check PRs run: | - { [[ $(git remote get-url origin) == *nf-core/atacseq ]] && [[ ${GITHUB_HEAD_REF} = "dev" ]]; } || [[ ${GITHUB_HEAD_REF} == "patch" ]] + { [[ $(git remote get-url origin) == *atacseq ]] && [[ ${GITHUB_HEAD_REF} = "dev" ]]; } || [[ ${GITHUB_HEAD_REF} == "patch" ]] From 1f586026cb911b626f04279c9b595c4cd57abc46 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 17 Dec 2019 10:53:25 +0000 Subject: [PATCH 067/226] Fix branch name --- .github/workflows/branch.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/.github/workflows/branch.yml b/.github/workflows/branch.yml index 37a3aa74..9ba92076 100755 --- a/.github/workflows/branch.yml +++ b/.github/workflows/branch.yml @@ -13,4 +13,4 @@ jobs: # PRs are only ok if coming from an nf-core `dev` branch or a fork `patch` branch - name: Check PRs run: | - { [[ $(git remote get-url origin) == *atacseq ]] && [[ ${GITHUB_HEAD_REF} = "dev" ]]; } || [[ ${GITHUB_HEAD_REF} == "patch" ]] + { [[ $(git remote get-url origin) == *nf-core/atacseq ]] && [[ ${GITHUB_HEAD_REF} = "dev" ]]; } || [[ ${GITHUB_HEAD_REF} == "patch" ]] From 8cdde6e6e616340bbc5d2d746ed5a0aa106ee6a2 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 17 Dec 2019 10:53:47 +0000 Subject: [PATCH 068/226] Export NXF_ANSI_LOG instead --- .github/workflows/ci.yml | 3 ++- 1 file changed, 2 insertions(+), 1 deletion(-) diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml index 39fc6c84..fa664e06 100755 --- a/.github/workflows/ci.yml +++ b/.github/workflows/ci.yml @@ -15,6 +15,7 @@ jobs: - name: Install Nextflow run: | export NXF_VER=${{ matrix.nxf_ver }} + export NXF_ANSI_LOG=false wget -qO- get.nextflow.io | bash sudo mv nextflow /usr/local/bin/ - name: Pull image @@ -22,4 +23,4 @@ jobs: docker pull nfcore/atacseq:dev && docker tag nfcore/atacseq:dev nfcore/atacseq:dev - name: Run test run: | - nextflow run ${GITHUB_WORKSPACE} -ansi-log false -profile test,docker + nextflow run ${GITHUB_WORKSPACE} -profile test,docker From e4c97a56f01e7da79eee38b6449f332398935a47 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Mon, 20 Jan 2020 12:25:41 +0000 Subject: [PATCH 069/226] Specify -T for sort commands --- main.nf | 12 ++++++------ 1 file changed, 6 insertions(+), 6 deletions(-) diff --git a/main.nf b/main.nf index bb05a290..5ebd65e5 100644 --- a/main.nf +++ b/main.nf @@ -966,7 +966,7 @@ process MergedLibBigWig { """ SCALE_FACTOR=\$(grep 'mapped (' $flagstat | awk '{print 1000000/\$1}') echo \$SCALE_FACTOR > ${prefix}.scale_factor.txt - genomeCoverageBed -ibam ${bam[0]} -bg -scale \$SCALE_FACTOR $pe_fragment $extend | sort -k1,1 -k2,2n > ${prefix}.bedGraph + genomeCoverageBed -ibam ${bam[0]} -bg -scale \$SCALE_FACTOR $pe_fragment $extend | sort -T '.' -k1,1 -k2,2n > ${prefix}.bedGraph bedGraphToBigWig ${prefix}.bedGraph $sizes ${prefix}.bigWig @@ -1213,7 +1213,7 @@ process MergedLibConsensusPeakSet { collapsecols = params.narrow_peak ? (["collapse"]*9).join(',') : (["collapse"]*8).join(',') expandparam = params.narrow_peak ? "--is_narrow_peak" : "" """ - sort -k1,1 -k2,2n ${peaks.collect{it.toString()}.sort().join(' ')} \\ + sort -T '.' -k1,1 -k2,2n ${peaks.collect{it.toString()}.sort().join(' ')} \\ | mergeBed -c $mergecols -o $collapsecols > ${prefix}.txt macs2_merged_expand.py \\ @@ -1265,7 +1265,7 @@ process MergedLibConsensusPeakSetAnnotate { -cpu $task.cpus \\ > ${prefix}.annotatePeaks.txt - cut -f2- ${prefix}.annotatePeaks.txt | awk 'NR==1; NR > 1 {print \$0 | "sort -k1,1 -k2,2n"}' | cut -f6- > tmp.txt + cut -f2- ${prefix}.annotatePeaks.txt | awk 'NR==1; NR > 1 {print \$0 | "sort -T '.' -k1,1 -k2,2n"}' | cut -f6- > tmp.txt paste $bool tmp.txt > ${prefix}.boolean.annotatePeaks.txt """ } @@ -1517,7 +1517,7 @@ process MergedRepBigWig { """ SCALE_FACTOR=\$(grep 'mapped (' $flagstat | awk '{print 1000000/\$1}') echo \$SCALE_FACTOR > ${prefix}.scale_factor.txt - genomeCoverageBed -ibam ${bam[0]} -bg -scale \$SCALE_FACTOR $pe_fragment $extend | sort -k1,1 -k2,2n > ${prefix}.bedGraph + genomeCoverageBed -ibam ${bam[0]} -bg -scale \$SCALE_FACTOR $pe_fragment $extend | sort -T '.' -k1,1 -k2,2n > ${prefix}.bedGraph bedGraphToBigWig ${prefix}.bedGraph $sizes ${prefix}.bigWig @@ -1680,7 +1680,7 @@ process MergedRepConsensusPeakSet { collapsecols = params.narrow_peak ? (["collapse"]*9).join(',') : (["collapse"]*8).join(',') expandparam = params.narrow_peak ? "--is_narrow_peak" : "" """ - sort -k1,1 -k2,2n ${peaks.collect{it.toString()}.sort().join(' ')} \\ + sort -T '.' -k1,1 -k2,2n ${peaks.collect{it.toString()}.sort().join(' ')} \\ | mergeBed -c $mergecols -o $collapsecols > ${prefix}.txt macs2_merged_expand.py \\ @@ -1731,7 +1731,7 @@ process MergedRepConsensusPeakSetAnnotate { -cpu $task.cpus \\ > ${prefix}.annotatePeaks.txt - cut -f2- ${prefix}.annotatePeaks.txt | awk 'NR==1; NR > 1 {print \$0 | "sort -k1,1 -k2,2n"}' | cut -f6- > tmp.txt + cut -f2- ${prefix}.annotatePeaks.txt | awk 'NR==1; NR > 1 {print \$0 | "sort -T '.' -k1,1 -k2,2n"}' | cut -f6- > tmp.txt paste $bool tmp.txt > ${prefix}.boolean.annotatePeaks.txt """ } From 38fb92ff0f16f281cf5c6e1dcae27a7dfe614fe9 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Mon, 20 Jan 2020 12:27:52 +0000 Subject: [PATCH 070/226] Update CHANGELOG --- CHANGELOG.md | 1 + 1 file changed, 1 insertion(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index b2e48240..04845097 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -15,6 +15,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Fixed` * [#118](https://github.com/nf-core/chipseq/issues/118) - Running on with SGE +* [#132][https://github.com/nf-core/chipseq/issues/132] - BigWig Error: sort: cannot create temporary file in '': Read-only file system * Make executables in `bin/` compatible with Python 3 ### `Dependencies` From e7ea781fc2b8299ad094d1d16dc96e54a07f89f9 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Mon, 20 Jan 2020 12:34:33 +0000 Subject: [PATCH 071/226] Fix markdownlint --- CHANGELOG.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 04845097..9824a3c8 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -15,7 +15,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Fixed` * [#118](https://github.com/nf-core/chipseq/issues/118) - Running on with SGE -* [#132][https://github.com/nf-core/chipseq/issues/132] - BigWig Error: sort: cannot create temporary file in '': Read-only file system +* [#132](https://github.com/nf-core/chipseq/issues/132) - BigWig Error: sort: cannot create temporary file in '': Read-only file system * Make executables in `bin/` compatible with Python 3 ### `Dependencies` From 3d5fb8a2ec6f6bfbc4aa0f06fe93862b7c68431f Mon Sep 17 00:00:00 2001 From: drpatelh Date: Mon, 20 Jan 2020 18:25:03 +0000 Subject: [PATCH 072/226] Fix missing annotation entries --- bin/plot_homer_annotatepeaks.r | 17 +++++++++++++---- 1 file changed, 13 insertions(+), 4 deletions(-) diff --git a/bin/plot_homer_annotatepeaks.r b/bin/plot_homer_annotatepeaks.r index 9865357c..57ca4b67 100755 --- a/bin/plot_homer_annotatepeaks.r +++ b/bin/plot_homer_annotatepeaks.r @@ -59,10 +59,19 @@ for (idx in 1:length(HomerFiles)) { sampleid = SampleIDs[idx] anno.dat <- read.table(HomerFiles[idx], sep="\t", header=TRUE,quote="") anno.dat <- anno.dat[,c("Annotation","Distance.to.TSS","Nearest.PromoterID")] - anno.dat <- anno.dat[which(!is.na(anno.dat$Distance.to.TSS)),] - if (nrow(anno.dat) == 0) { - quit(save = "no", status = 0, runLast = FALSE) - } + + ## REPLACE UNASSIGNED FEATURE ENTRIES WITH SENSIBLE VALUES + unassigned <- which(is.na(as.character(anno.dat$Distance.to.TSS))) + anno.dat$Distance.to.TSS[unassigned] <- 1000000 + + anno.dat$Annotation <- as.character(anno.dat$Annotation) + anno.dat$Annotation[unassigned] <- "Unassigned" + anno.dat$Annotation <- as.factor(anno.dat$Annotation) + + anno.dat$Nearest.PromoterID <- as.character(anno.dat$Nearest.PromoterID) + anno.dat$Nearest.PromoterID[unassigned] <- "Unassigned" + anno.dat$Nearest.PromoterID <- as.factor(anno.dat$Nearest.PromoterID) + anno.dat$name <- rep(sampleid,nrow(anno.dat)) anno.dat$Distance.to.TSS <- abs(anno.dat$Distance.to.TSS) + 1 plot.dat <- rbind(plot.dat,anno.dat) From a3f7f51d6171d1a943dc5237216635abfaebe0cf Mon Sep 17 00:00:00 2001 From: drpatelh Date: Mon, 20 Jan 2020 18:28:15 +0000 Subject: [PATCH 073/226] Update CHANGELOG --- CHANGELOG.md | 1 + 1 file changed, 1 insertion(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index 9824a3c8..c8d88d88 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -16,6 +16,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * [#118](https://github.com/nf-core/chipseq/issues/118) - Running on with SGE * [#132](https://github.com/nf-core/chipseq/issues/132) - BigWig Error: sort: cannot create temporary file in '': Read-only file system +* [#73](https://github.com/nf-core/atacseq/issues/73) - macs_annotatePeaks.mLb.clN.summary.txt file is not created * Make executables in `bin/` compatible with Python 3 ### `Dependencies` From d65bad41ac513555bb3dd2f661877b2286db6b4a Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 29 Jan 2020 14:34:07 +0000 Subject: [PATCH 074/226] Remove .travis.yml --- .travis.yml | 48 ------------------------------------------------ 1 file changed, 48 deletions(-) delete mode 100755 .travis.yml diff --git a/.travis.yml b/.travis.yml deleted file mode 100755 index 3b07ff58..00000000 --- a/.travis.yml +++ /dev/null @@ -1,48 +0,0 @@ -sudo: required -language: python -jdk: openjdk8 -services: docker -python: '3.6' -cache: pip -matrix: - fast_finish: true - -before_install: - # PRs to master are only ok if coming from dev branch - #- '[ $TRAVIS_PULL_REQUEST = "false" ] || [ $TRAVIS_BRANCH != "master" ] || ([ $TRAVIS_PULL_REQUEST_SLUG = $TRAVIS_REPO_SLUG ] && [ $TRAVIS_PULL_REQUEST_BRANCH = "dev" ]) || [ $TRAVIS_PULL_REQUEST_BRANCH = "patch" ]' - - '[ $TRAVIS_PULL_REQUEST = "false" ] || [ $TRAVIS_BRANCH != "master" ] || ([ $TRAVIS_PULL_REQUEST_SLUG = $TRAVIS_REPO_SLUG ] && ([ $TRAVIS_PULL_REQUEST_BRANCH = "dev" ] || [ $TRAVIS_PULL_REQUEST_BRANCH = "patch" ]))' - # Pull the docker image first so the test doesn't wait for this - - docker pull nfcore/atacseq:dev - # Fake the tag locally so that the pipeline runs properly - # Looks weird when this is :dev to :dev, but makes sense when testing code for a release (:dev to :1.0.1) - - docker tag nfcore/atacseq:dev nfcore/atacseq:dev - -install: - # Install Nextflow - - mkdir /tmp/nextflow && cd /tmp/nextflow - - wget -qO- get.nextflow.io | bash - - sudo ln -s /tmp/nextflow/nextflow /usr/local/bin/nextflow - # Install nf-core/tools - - pip install --upgrade pip - - pip install nf-core - # Reset - - mkdir ${TRAVIS_BUILD_DIR}/tests && cd ${TRAVIS_BUILD_DIR}/tests - # Install markdownlint-cli - - sudo apt-get install npm && npm install -g markdownlint-cli - -env: - # Tower token is to inspect runs on https://tower.nf - # Use public mailbox nf-core@mailinator.com to log in: https://www.mailinator.com/v3/index.jsp?zone=public&query=nf-core - # Specify a minimum NF version that should be tested and work - - NXF_VER='19.10.0' TOWER_ACCESS_TOKEN="1c1f493bc2703472d6f1b9f6fb9e9d117abab7b1" - # Plus: get the latest NF version and check that it works - - NXF_VER='' TOWER_ACCESS_TOKEN="1c1f493bc2703472d6f1b9f6fb9e9d117abab7b1" - - -script: - # Lint the pipeline code - - nf-core lint ${TRAVIS_BUILD_DIR} - # Lint the documentation - - markdownlint ${TRAVIS_BUILD_DIR} -c ${TRAVIS_BUILD_DIR}/.github/markdownlint.yml - # Run the pipeline with the test profile - - nextflow run ${TRAVIS_BUILD_DIR} -profile test,docker -ansi-log false -name atacseq-${TRAVIS_EVENT_TYPE}-${TRAVIS_PULL_REQUEST}-${TRAVIS_COMMIT:0:6}-test-description From 8b0f863f21f4f907bf69bf7184fdde8a0fb3f005 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 29 Jan 2020 14:34:18 +0000 Subject: [PATCH 075/226] Update nf-core/base --- Dockerfile | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/Dockerfile b/Dockerfile index abb90542..9e121402 100755 --- a/Dockerfile +++ b/Dockerfile @@ -1,4 +1,4 @@ -FROM nfcore/base:1.7 +FROM nfcore/base:1.8 LABEL authors="Harshil Patel" \ description="Docker image containing all software requirements for the nf-core/atacseq pipeline" From 9526922287a2173df6df617931b1329c0eaf25ee Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 29 Jan 2020 17:12:27 +0000 Subject: [PATCH 076/226] Add readme path to igenomes.config --- conf/igenomes.config | 23 +++++++++++++++++++++++ 1 file changed, 23 insertions(+) diff --git a/conf/igenomes.config b/conf/igenomes.config index 37217cf4..2de92422 100755 --- a/conf/igenomes.config +++ b/conf/igenomes.config @@ -18,6 +18,7 @@ params { bismark = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Annotation/README.txt" mito_name = "MT" macs_gsize = "2.7e9" blacklist = "${baseDir}/assets/blacklists/GRCh37-blacklist.bed" @@ -42,6 +43,7 @@ params { bismark = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Annotation/README.txt" mito_name = "MT" macs_gsize = "1.87e9" blacklist = "${baseDir}/assets/blacklists/GRCm38-blacklist.bed" @@ -54,6 +56,7 @@ params { bismark = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Annotation/README.txt" mito_name = "Mt" } 'EB2' { @@ -64,6 +67,7 @@ params { bismark = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Annotation/README.txt" } 'UMD3.1' { fasta = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/WholeGenomeFasta/genome.fa" @@ -73,6 +77,7 @@ params { bismark = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Annotation/README.txt" mito_name = "MT" } 'WBcel235' { @@ -94,6 +99,7 @@ params { bismark = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Annotation/README.txt" mito_name = "MT" } 'GRCz10' { @@ -125,6 +131,7 @@ params { bismark = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Annotation/README.txt" mito_name = "MT" } 'EB1' { @@ -135,6 +142,7 @@ params { bismark = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Annotation/README.txt" } 'Galgal4' { fasta = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/WholeGenomeFasta/genome.fa" @@ -154,6 +162,7 @@ params { bismark = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Annotation/README.txt" } 'Mmul_1' { fasta = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/WholeGenomeFasta/genome.fa" @@ -163,6 +172,7 @@ params { bismark = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Annotation/README.txt" mito_name = "MT" } 'IRGSP-1.0' { @@ -183,6 +193,7 @@ params { bismark = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Annotation/README.txt" mito_name = "MT" } 'Rnor_6.0' { @@ -214,6 +225,7 @@ params { bismark = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Annotation/README.txt" mito_name = "MT" macs_gsize = "1.21e7" } @@ -225,6 +237,7 @@ params { bismark = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Annotation/README.txt" } 'Sscrofa10.2' { fasta = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/WholeGenomeFasta/genome.fa" @@ -234,6 +247,7 @@ params { bismark = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Annotation/README.txt" mito_name = "MT" } 'AGPv3' { @@ -266,6 +280,7 @@ params { bismark = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Annotation/README.txt" mito_name = "chrM" macs_gsize = "2.7e9" blacklist = "${baseDir}/assets/blacklists/hg19-blacklist.bed" @@ -278,6 +293,7 @@ params { bismark = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Annotation/README.txt" mito_name = "chrM" macs_gsize = "1.87e9" blacklist = "${baseDir}/assets/blacklists/mm10-blacklist.bed" @@ -300,6 +316,7 @@ params { bismark = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Annotation/README.txt" mito_name = "chrM" macs_gsize = "9e7" } @@ -311,6 +328,7 @@ params { bismark = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Annotation/README.txt" mito_name = "chrM" } 'danRer10' { @@ -342,6 +360,7 @@ params { bismark = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Annotation/README.txt" mito_name = "chrM" } 'galGal4' { @@ -352,6 +371,7 @@ params { bismark = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Annotation/README.txt" mito_name = "chrM" } 'panTro4' { @@ -362,6 +382,7 @@ params { bismark = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Annotation/README.txt" mito_name = "chrM" } 'rn6' { @@ -380,6 +401,7 @@ params { bowtie2 = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/Bowtie2Index/" star = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/STARIndex/" bismark = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/BismarkIndex/" + readme = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Annotation/README.txt" mito_name = "chrM" macs_gsize = "1.2e7" } @@ -391,6 +413,7 @@ params { bismark = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Annotation/README.txt" mito_name = "chrM" } } From 803501a0c0b72b1fc27591d512b6933a1b2fb4ea Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 29 Jan 2020 17:13:46 +0000 Subject: [PATCH 077/226] Add igenomes readme logic --- main.nf | 13 ++++++++----- 1 file changed, 8 insertions(+), 5 deletions(-) diff --git a/main.nf b/main.nf index 5ebd65e5..124bb29b 100644 --- a/main.nf +++ b/main.nf @@ -123,6 +123,7 @@ params.gene_bed = params.genome ? params.genomes[ params.genome ].bed12 ?: false params.mito_name = params.genome ? params.genomes[ params.genome ].mito_name ?: false : false params.macs_gsize = params.genome ? params.genomes[ params.genome ].macs_gsize ?: false : false params.blacklist = params.genome ? params.genomes[ params.genome ].blacklist ?: false : false +params.anno_readme = params.genome ? params.genomes[ params.genome ].readme ?: false : false // Global variables def PEAK_TYPE = params.narrow_peak ? "narrowPeak" : "broadPeak" @@ -167,11 +168,12 @@ ch_mrep_deseq2_clustering_header = file("$baseDir/assets/multiqc/mrep_deseq2_clu //////////////////////////////////////////////////// // Validate inputs -if (params.input) { ch_input = file(params.input, checkIfExists: true) } else { exit 1, "Samples design file not specified!" } -if (params.gtf) { ch_gtf = file(params.gtf, checkIfExists: true) } else { exit 1, "GTF annotation file not specified!" } -if (params.gene_bed) { ch_gene_bed = file(params.gene_bed, checkIfExists: true) } -if (params.tss_bed) { ch_tss_bed = file(params.tss_bed, checkIfExists: true) } -if (params.blacklist) { ch_blacklist = Channel.fromPath(params.blacklist, checkIfExists: true) } else { ch_blacklist = Channel.empty() } +if (params.input) { ch_input = file(params.input, checkIfExists: true) } else { exit 1, "Samples design file not specified!" } +if (params.gtf) { ch_gtf = file(params.gtf, checkIfExists: true) } else { exit 1, "GTF annotation file not specified!" } +if (params.gene_bed) { ch_gene_bed = file(params.gene_bed, checkIfExists: true) } +if (params.tss_bed) { ch_tss_bed = file(params.tss_bed, checkIfExists: true) } +if (params.blacklist) { ch_blacklist = Channel.fromPath(params.blacklist, checkIfExists: true) } else { ch_blacklist = Channel.empty() } +if (params.anno_readme) { ch_anno_readme = Channel.fromPath(params.anno_readme) } else { ch_anno_readme = Channel.empty() } if (params.fasta) { lastPath = params.fasta.lastIndexOf(File.separator) @@ -1906,6 +1908,7 @@ process MultiQC { input: file multiqc_config from ch_multiqc_config + file readme from ch_anno_readme.collect().ifEmpty([]) file ('software_versions/*') from ch_software_versions_mqc.collect() file ('workflow_summary/*') from create_workflow_summary(summary) From 7e2fe07e1f2c8e3dc3b87b711364b622804ce80d Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 30 Jan 2020 13:45:17 +0000 Subject: [PATCH 078/226] Implement anno README --- main.nf | 3261 ++++++++++++++++++++++++++++--------------------------- 1 file changed, 1631 insertions(+), 1630 deletions(-) diff --git a/main.nf b/main.nf index 124bb29b..fc7dd155 100644 --- a/main.nf +++ b/main.nf @@ -173,7 +173,7 @@ if (params.gtf) { ch_gtf = file(params.gtf, checkIfExists: true) } else if (params.gene_bed) { ch_gene_bed = file(params.gene_bed, checkIfExists: true) } if (params.tss_bed) { ch_tss_bed = file(params.tss_bed, checkIfExists: true) } if (params.blacklist) { ch_blacklist = Channel.fromPath(params.blacklist, checkIfExists: true) } else { ch_blacklist = Channel.empty() } -if (params.anno_readme) { ch_anno_readme = Channel.fromPath(params.anno_readme) } else { ch_anno_readme = Channel.empty() } +if (params.anno_readme && file(params.anno_readme).exists()) { ch_anno_readme = Channel.fromPath(params.anno_readme) } else { ch_anno_readme = Channel.empty() } if (params.fasta) { lastPath = params.fasta.lastIndexOf(File.separator) @@ -446,10 +446,12 @@ process MakeGenomeFilter { input: file fasta from ch_fasta file blacklist from ch_blacklist.ifEmpty([]) + file readme from ch_anno_readme.ifEmpty([]) output: - file "$fasta" into ch_genome_fasta // FASTA FILE FOR IGV - file "*.fai" into ch_genome_fai // FAI INDEX FOR REFERENCE GENOME + file "$fasta" // FASTA FILE FOR IGV + file "$readme" // AWS IGENOMES FILE CONTAINING ANNOTATION VERSION + file "*.fai" // FAI INDEX FOR REFERENCE GENOME file "*.bed" into ch_genome_filter_regions // BED FILE WITHOUT BLACKLIST REGIONS & MITOCHONDRIAL CONTIG FOR FILTERING file "*.txt" into ch_genome_autosomes // TEXT FILE CONTAINING LISTING OF AUTOSOMAL CHROMOSOMES FOR ATAQV file "*.sizes" into ch_genome_sizes_mlib_bigwig, // CHROMOSOME SIZES FILE FOR BEDTOOLS @@ -467,1633 +469,1632 @@ process MakeGenomeFilter { """ } -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// -/* -- -- */ -/* -- FASTQ QC -- */ -/* -- -- */ -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// - -/* - * STEP 1 - FastQC - */ -process FastQC { - tag "$name" - label 'process_medium' - publishDir "${params.outdir}/fastqc", mode: 'copy', - saveAs: { filename -> - filename.endsWith(".zip") ? "zips/$filename" : "$filename" - } - - when: - !params.skip_fastqc - - input: - set val(name), file(reads) from ch_raw_reads_fastqc - - output: - file "*.{zip,html}" into ch_fastqc_reports_mqc - - script: - // Added soft-links to original fastqs for consistent naming in MultiQC - if (params.single_end) { - """ - [ ! -f ${name}.fastq.gz ] && ln -s $reads ${name}.fastq.gz - fastqc -q -t $task.cpus ${name}.fastq.gz - """ - } else { - """ - [ ! -f ${name}_1.fastq.gz ] && ln -s ${reads[0]} ${name}_1.fastq.gz - [ ! -f ${name}_2.fastq.gz ] && ln -s ${reads[1]} ${name}_2.fastq.gz - fastqc -q -t $task.cpus ${name}_1.fastq.gz - fastqc -q -t $task.cpus ${name}_2.fastq.gz - """ - } -} - -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// -/* -- -- */ -/* -- ADAPTER TRIMMING -- */ -/* -- -- */ -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// - -/* - * STEP 2 - Trim Galore! - */ -if (params.skip_trimming) { - ch_trimmed_reads = ch_raw_reads_trimgalore - ch_trimgalore_results_mqc = Channel.empty() - ch_trimgalore_fastqc_reports_mqc = Channel.empty() -} else { - process TrimGalore { - tag "$name" - label 'process_high' - publishDir "${params.outdir}/trim_galore", mode: 'copy', - saveAs: { filename -> - if (filename.endsWith(".html")) "fastqc/$filename" - else if (filename.endsWith(".zip")) "fastqc/zips/$filename" - else if (filename.endsWith("trimming_report.txt")) "logs/$filename" - else params.save_trimmed ? filename : null - } - - input: - set val(name), file(reads) from ch_raw_reads_trimgalore - - output: - set val(name), file("*.fq.gz") into ch_trimmed_reads - file "*.txt" into ch_trimgalore_results_mqc - file "*.{zip,html}" into ch_trimgalore_fastqc_reports_mqc - - script: - // Calculate number of --cores for TrimGalore based on value of task.cpus - // See: https://github.com/FelixKrueger/TrimGalore/blob/master/Changelog.md#version-060-release-on-1-mar-2019 - // See: https://github.com/nf-core/atacseq/pull/65 - def cores = 1 - if (task.cpus) { - cores = (task.cpus as int) - 4 - if (params.single_end) cores = (task.cpus as int) - 3 - if (cores < 1) cores = 1 - if (cores > 4) cores = 4 - } - - // Added soft-links to original fastqs for consistent naming in MultiQC - c_r1 = params.clip_r1 > 0 ? "--clip_r1 ${params.clip_r1}" : '' - c_r2 = params.clip_r2 > 0 ? "--clip_r2 ${params.clip_r2}" : '' - tpc_r1 = params.three_prime_clip_r1 > 0 ? "--three_prime_clip_r1 ${params.three_prime_clip_r1}" : '' - tpc_r2 = params.three_prime_clip_r2 > 0 ? "--three_prime_clip_r2 ${params.three_prime_clip_r2}" : '' - nextseq = params.trim_nextseq > 0 ? "--nextseq ${params.trim_nextseq}" : '' - - // Added soft-links to original fastqs for consistent naming in MultiQC - if (params.single_end) { - """ - [ ! -f ${name}.fastq.gz ] && ln -s $reads ${name}.fastq.gz - trim_galore --cores $cores --fastqc --gzip $c_r1 $tpc_r1 $nextseq ${name}.fastq.gz - """ - } else { - """ - [ ! -f ${name}_1.fastq.gz ] && ln -s ${reads[0]} ${name}_1.fastq.gz - [ ! -f ${name}_2.fastq.gz ] && ln -s ${reads[1]} ${name}_2.fastq.gz - trim_galore --cores $cores --paired --fastqc --gzip $c_r1 $c_r2 $tpc_r1 $tpc_r2 $nextseq ${name}_1.fastq.gz ${name}_2.fastq.gz - """ - } - } -} - -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// -/* -- -- */ -/* -- ALIGN -- */ -/* -- -- */ -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// - -/* - * STEP 3.1 - Align read 1 with bwa - */ -process BWAMem { - tag "$name" - label 'process_high' - - input: - set val(name), file(reads) from ch_trimmed_reads - file index from ch_bwa_index.collect() - - output: - set val(name), file("*.bam") into ch_bwa_bam - - script: - prefix = "${name}.Lb" - rg = "\'@RG\\tID:${name}\\tSM:${name.split('_')[0..-2].join('_')}\\tPL:ILLUMINA\\tLB:${name}\\tPU:1\'" - if (params.seq_center) { - rg = "\'@RG\\tID:${name}\\tSM:${name.split('_')[0..-2].join('_')}\\tPL:ILLUMINA\\tLB:${name}\\tPU:1\\tCN:${params.seq_center}\'" - } - """ - bwa mem \\ - -t $task.cpus \\ - -M \\ - -R $rg \\ - ${index}/${bwa_base} \\ - $reads \\ - | samtools view -@ $task.cpus -b -h -F 0x0100 -O BAM -o ${prefix}.bam - - """ -} - -/* - * STEP 3.2 - Convert .bam to coordinate sorted .bam - */ -process SortBAM { - tag "$name" - label 'process_medium' - if (params.save_align_intermeds) { - publishDir path: "${params.outdir}/bwa/library", mode: 'copy', - saveAs: { filename -> - if (filename.endsWith(".flagstat")) "samtools_stats/$filename" - else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" - else if (filename.endsWith(".stats")) "samtools_stats/$filename" - else filename - } - } - - input: - set val(name), file(bam) from ch_bwa_bam - - output: - set val(name), file("*.sorted.{bam,bam.bai}") into ch_sort_bam_merge - file "*.{flagstat,idxstats,stats}" into ch_sort_bam_flagstat_mqc - - script: - prefix = "${name}.Lb" - """ - samtools sort -@ $task.cpus -o ${prefix}.sorted.bam -T $name $bam - samtools index ${prefix}.sorted.bam - samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat - samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats - samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats - """ -} - -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// -/* -- -- */ -/* -- MERGE LIBRARY BAM -- */ -/* -- -- */ -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// - -/* - * STEP 4.1 Merge BAM files for all libraries from same replicate - */ -ch_sort_bam_merge - .map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] } - .groupTuple(by: [0]) - .map { it -> [ it[0], it[1].flatten() ] } - .set { ch_sort_bam_merge } - -process MergedLibBAM { - tag "$name" - label 'process_medium' - publishDir "${params.outdir}/bwa/mergedLibrary", mode: 'copy', - saveAs: { filename -> - if (filename.endsWith(".flagstat")) "samtools_stats/$filename" - else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" - else if (filename.endsWith(".stats")) "samtools_stats/$filename" - else if (filename.endsWith(".metrics.txt")) "picard_metrics/$filename" - else params.save_align_intermeds ? filename : null - } - - input: - set val(name), file(bams) from ch_sort_bam_merge - - output: - set val(name), file("*${prefix}.sorted.{bam,bam.bai}") into ch_mlib_bam_filter, - ch_mlib_bam_preseq, - ch_mlib_bam_ataqv - file "*.{flagstat,idxstats,stats}" into ch_mlib_bam_stats_mqc - file "*.txt" into ch_mlib_bam_metrics_mqc - - script: - prefix = "${name}.mLb.mkD" - bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() - def avail_mem = 3 - if (!task.memory) { - log.info "[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." - } else { - avail_mem = task.memory.toGiga() - } - if (bam_files.size() > 1) { - """ - picard -Xmx${avail_mem}g MergeSamFiles \\ - ${'INPUT='+bam_files.join(' INPUT=')} \\ - OUTPUT=${name}.sorted.bam \\ - SORT_ORDER=coordinate \\ - VALIDATION_STRINGENCY=LENIENT \\ - TMP_DIR=tmp - samtools index ${name}.sorted.bam - - picard -Xmx${avail_mem}g MarkDuplicates \\ - INPUT=${name}.sorted.bam \\ - OUTPUT=${prefix}.sorted.bam \\ - ASSUME_SORTED=true \\ - REMOVE_DUPLICATES=false \\ - METRICS_FILE=${prefix}.MarkDuplicates.metrics.txt \\ - VALIDATION_STRINGENCY=LENIENT \\ - TMP_DIR=tmp - - samtools index ${prefix}.sorted.bam - samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats - samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat - samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats - """ - } else { - """ - picard -Xmx${avail_mem}g MarkDuplicates \\ - INPUT=${bam_files[0]} \\ - OUTPUT=${prefix}.sorted.bam \\ - ASSUME_SORTED=true \\ - REMOVE_DUPLICATES=false \\ - METRICS_FILE=${prefix}.MarkDuplicates.metrics.txt \\ - VALIDATION_STRINGENCY=LENIENT \\ - TMP_DIR=tmp - - samtools index ${prefix}.sorted.bam - samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats - samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat - samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats - """ - } -} - -/* - * STEP 4.2 Filter BAM file at merged library-level - */ -process MergedLibBAMFilter { - tag "$name" - label 'process_medium' - publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', - saveAs: { filename -> - if (params.single_end || params.save_align_intermeds) { - if (filename.endsWith(".flagstat")) "samtools_stats/$filename" - else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" - else if (filename.endsWith(".stats")) "samtools_stats/$filename" - else if (filename.endsWith(".sorted.bam")) filename - else if (filename.endsWith(".sorted.bam.bai")) filename - else null - } - } - - input: - set val(name), file(bam) from ch_mlib_bam_filter - file bed from ch_genome_filter_regions.collect() - file bamtools_filter_config from ch_bamtools_filter_config - - output: - set val(name), file("*.{bam,bam.bai}") into ch_mlib_filter_bam - set val(name), file("*.flagstat") into ch_mlib_filter_bam_flagstat - file "*.{idxstats,stats}" into ch_mlib_filter_bam_stats_mqc - - script: - prefix = params.single_end ? "${name}.mLb.clN" : "${name}.mLb.flT" - filter_params = params.single_end ? "-F 0x004" : "-F 0x004 -F 0x0008 -f 0x001" - dup_params = params.keep_dups ? "" : "-F 0x0400" - multimap_params = params.keep_multi_map ? "" : "-q 1" - blacklist_params = params.blacklist ? "-L $bed" : "" - name_sort_bam = params.single_end ? "" : "samtools sort -n -@ $task.cpus -o ${prefix}.bam -T $prefix ${prefix}.sorted.bam" - """ - samtools view \\ - $filter_params \\ - $dup_params \\ - $multimap_params \\ - $blacklist_params \\ - -b ${bam[0]} \\ - | bamtools filter \\ - -out ${prefix}.sorted.bam \\ - -script $bamtools_filter_config - - samtools index ${prefix}.sorted.bam - samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat - samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats - samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats - - $name_sort_bam - """ -} - -/* - * STEP 4.3 Remove orphan reads from paired-end BAM file - */ -if (params.single_end) { - ch_mlib_filter_bam - .into { ch_mlib_rm_orphan_bam_metrics; - ch_mlib_rm_orphan_bam_bigwig; - ch_mlib_rm_orphan_bam_macs; - ch_mlib_rm_orphan_bam_plotfingerprint; - ch_mlib_rm_orphan_bam_mrep; - ch_mlib_name_bam_mlib_counts; - ch_mlib_name_bam_mrep_counts } - - ch_mlib_filter_bam_flagstat - .into { ch_mlib_rm_orphan_flagstat_bigwig; - ch_mlib_rm_orphan_flagstat_macs; - ch_mlib_rm_orphan_flagstat_mqc } - - ch_mlib_filter_bam_stats_mqc - .set { ch_mlib_rm_orphan_stats_mqc } -} else { - process MergedLibBAMRemoveOrphan { - tag "$name" - label 'process_medium' - publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', - saveAs: { filename -> - if (filename.endsWith(".flagstat")) "samtools_stats/$filename" - else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" - else if (filename.endsWith(".stats")) "samtools_stats/$filename" - else if (filename.endsWith(".sorted.bam")) filename - else if (filename.endsWith(".sorted.bam.bai")) filename - else null - } - - input: - set val(name), file(bam) from ch_mlib_filter_bam - - output: - set val(name), file("*.sorted.{bam,bam.bai}") into ch_mlib_rm_orphan_bam_metrics, - ch_mlib_rm_orphan_bam_bigwig, - ch_mlib_rm_orphan_bam_macs, - ch_mlib_rm_orphan_bam_plotfingerprint, - ch_mlib_rm_orphan_bam_mrep - set val(name), file("${prefix}.bam") into ch_mlib_name_bam_mlib_counts, - ch_mlib_name_bam_mrep_counts - set val(name), file("*.flagstat") into ch_mlib_rm_orphan_flagstat_bigwig, - ch_mlib_rm_orphan_flagstat_macs, - ch_mlib_rm_orphan_flagstat_mqc - file "*.{idxstats,stats}" into ch_mlib_rm_orphan_stats_mqc - - script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ - prefix = "${name}.mLb.clN" - """ - bampe_rm_orphan.py ${bam[0]} ${prefix}.bam --only_fr_pairs - - samtools sort -@ $task.cpus -o ${prefix}.sorted.bam -T $prefix ${prefix}.bam - samtools index ${prefix}.sorted.bam - samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat - samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats - samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats - """ - } -} - -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// -/* -- -- */ -/* -- MERGE LIBRARY BAM POST-ANALYSIS -- */ -/* -- -- */ -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// - -/* - * STEP 5.1 preseq analysis after merging libraries and before filtering - */ -process MergedLibPreseq { - tag "$name" - label 'process_low' - publishDir "${params.outdir}/bwa/mergedLibrary/preseq", mode: 'copy' - - when: - !params.skip_preseq - - input: - set val(name), file(bam) from ch_mlib_bam_preseq - - output: - file "*.ccurve.txt" into ch_mlib_preseq_mqc - - script: - prefix = "${name}.mLb.mkD" - """ - preseq lc_extrap -v -output ${prefix}.ccurve.txt -bam ${bam[0]} - """ -} - -/* - * STEP 5.2 Picard CollectMultipleMetrics after merging libraries and filtering - */ -process MergedLibMetrics { - tag "$name" - label 'process_medium' - publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', - saveAs: { filename -> - if (filename.endsWith("_metrics")) "picard_metrics/$filename" - else if (filename.endsWith(".pdf")) "picard_metrics/pdf/$filename" - else null - } - - when: - !params.skip_picard_metrics - - input: - set val(name), file(bam) from ch_mlib_rm_orphan_bam_metrics - file fasta from ch_fasta - - output: - file "*_metrics" into ch_mlib_collectmetrics_mqc - file "*.pdf" into ch_mlib_collectmetrics_pdf - - script: - prefix = "${name}.mLb.clN" - def avail_mem = 3 - if (!task.memory) { - log.info "[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." - } else { - avail_mem = task.memory.toGiga() - } - """ - picard -Xmx${avail_mem}g CollectMultipleMetrics \\ - INPUT=${bam[0]} \\ - OUTPUT=${prefix}.CollectMultipleMetrics \\ - REFERENCE_SEQUENCE=$fasta \\ - VALIDATION_STRINGENCY=LENIENT \\ - TMP_DIR=tmp - """ -} - -/* - * STEP 5.3 Read depth normalised bigWig - */ -process MergedLibBigWig { - tag "$name" - label 'process_medium' - publishDir "${params.outdir}/bwa/mergedLibrary/bigwig", mode: 'copy', - saveAs: { filename -> - if (filename.endsWith("scale_factor.txt")) "scale/$filename" - else if (filename.endsWith(".bigWig")) "$filename" - else null - } - - input: - set val(name), file(bam), file(flagstat) from ch_mlib_rm_orphan_bam_bigwig.join(ch_mlib_rm_orphan_flagstat_bigwig, by: [0]) - file sizes from ch_genome_sizes_mlib_bigwig.collect() - - output: - set val(name), file("*.bigWig") into ch_mlib_bigwig_plotprofile - file "*scale_factor.txt" into ch_mlib_bigwig_scale - file "*igv.txt" into ch_mlib_bigwig_igv - - script: - prefix = "${name}.mLb.clN" - pe_fragment = params.single_end ? "" : "-pc" - extend = (params.single_end && params.fragment_size > 0) ? "-fs ${params.fragment_size}" : '' - """ - SCALE_FACTOR=\$(grep 'mapped (' $flagstat | awk '{print 1000000/\$1}') - echo \$SCALE_FACTOR > ${prefix}.scale_factor.txt - genomeCoverageBed -ibam ${bam[0]} -bg -scale \$SCALE_FACTOR $pe_fragment $extend | sort -T '.' -k1,1 -k2,2n > ${prefix}.bedGraph - - bedGraphToBigWig ${prefix}.bedGraph $sizes ${prefix}.bigWig - - find * -type f -name "*.bigWig" -exec echo -e "bwa/mergedLibrary/bigwig/"{}"\\t0,0,178" \\; > ${prefix}.bigWig.igv.txt - """ -} - -/* - * STEP 5.4 generate gene body coverage plot with deepTools - */ -process MergedLibPlotProfile { - tag "$name" - label 'process_high' - publishDir "${params.outdir}/bwa/mergedLibrary/deepTools/plotProfile", mode: 'copy' - - when: - !params.skip_plot_profile - - input: - set val(name), file(bigwig) from ch_mlib_bigwig_plotprofile - file bed from ch_gene_bed - - output: - file '*.{gz,pdf}' into ch_mlib_plotprofile_results - file '*.plotProfile.tab' into ch_mlib_plotprofile_mqc - - script: - prefix = "${name}.mLb.clN" - """ - computeMatrix scale-regions \\ - --regionsFileName $bed \\ - --scoreFileName $bigwig \\ - --outFileName ${prefix}.computeMatrix.mat.gz \\ - --outFileNameMatrix ${prefix}.computeMatrix.vals.mat.gz \\ - --regionBodyLength 1000 \\ - --beforeRegionStartLength 3000 \\ - --afterRegionStartLength 3000 \\ - --skipZeros \\ - --smartLabels \\ - --numberOfProcessors $task.cpus - - plotProfile --matrixFile ${prefix}.computeMatrix.mat.gz \\ - --outFileName ${prefix}.plotProfile.pdf \\ - --outFileNameData ${prefix}.plotProfile.tab - """ -} - -/* - * STEP 5.5 deepTools plotFingerprint - */ -process MergedLibPlotFingerprint { - tag "$name" - label 'process_high' - publishDir "${params.outdir}/bwa/mergedLibrary/deepTools/plotFingerprint", mode: 'copy' - - when: - !params.skip_plot_fingerprint - - input: - set val(name), file(bam) from ch_mlib_rm_orphan_bam_plotfingerprint - - output: - file '*.{txt,pdf}' into ch_mlib_plotfingerprint_results - file '*.raw.txt' into ch_mlib_plotfingerprint_mqc - - script: - prefix = "${name}.mLb.clN" - extend = (params.single_end && params.fragment_size > 0) ? "--extendReads ${params.fragment_size}" : '' - """ - plotFingerprint \\ - --bamfiles ${bam[0]} \\ - --plotFile ${prefix}.plotFingerprint.pdf \\ - $extend \\ - --labels $prefix \\ - --outRawCounts ${prefix}.plotFingerprint.raw.txt \\ - --outQualityMetrics ${prefix}.plotFingerprint.qcmetrics.txt \\ - --skipZeros \\ - --numberOfProcessors $task.cpus \\ - --numberOfSamples $params.fingerprint_bins - """ -} - -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// -/* -- -- */ -/* -- MERGE LIBRARY PEAK ANALYSIS -- */ -/* -- -- */ -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// - -/* - * STEP 6.1 Call peaks with MACS2 and calculate FRiP score - */ -process MergedLibMACSCallPeak { - tag "$name" - label 'process_medium' - publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}", mode: 'copy', - saveAs: { filename -> - if (filename.endsWith(".tsv")) "qc/$filename" - else if (filename.endsWith(".igv.txt")) null - else filename - } - - when: - params.macs_gsize - - input: - set val(name), file(bam), file(flagstat) from ch_mlib_rm_orphan_bam_macs.join(ch_mlib_rm_orphan_flagstat_macs, by: [0]) - file mlib_peak_count_header from ch_mlib_peak_count_header - file mlib_frip_score_header from ch_mlib_frip_score_header - - output: - set val(name), file("*.{bed,xls,gappedPeak,bdg}") into ch_mlib_macs_output - set val(name), file("*$PEAK_TYPE") into ch_mlib_macs_homer, - ch_mlib_macs_qc, - ch_mlib_macs_consensus, - ch_mlib_macs_ataqv - file "*igv.txt" into ch_mlib_macs_igv - file "*_mqc.tsv" into ch_mlib_macs_mqc - - script: - prefix = "${name}.mLb.clN" - broad = params.narrow_peak ? '' : "--broad --broad-cutoff ${params.broad_cutoff}" - format = params.single_end ? "BAM" : "BAMPE" - pileup = params.save_macs_pileup ? "-B --SPMR" : "" - """ - macs2 callpeak \\ - -t ${bam[0]} \\ - $broad \\ - -f $format \\ - -g $params.macs_gsize \\ - -n $prefix \\ - $pileup \\ - --keep-dup all \\ - --nomodel - - cat ${prefix}_peaks.${PEAK_TYPE} | wc -l | awk -v OFS='\t' '{ print "${name}", \$1 }' | cat $mlib_peak_count_header - > ${prefix}_peaks.count_mqc.tsv - - READS_IN_PEAKS=\$(intersectBed -a ${bam[0]} -b ${prefix}_peaks.${PEAK_TYPE} -bed -c -f 0.20 | awk -F '\t' '{sum += \$NF} END {print sum}') - grep 'mapped (' $flagstat | awk -v a="\$READS_IN_PEAKS" -v OFS='\t' '{print "${name}", a/\$1}' | cat $mlib_frip_score_header - > ${prefix}_peaks.FRiP_mqc.tsv - - find * -type f -name "*.${PEAK_TYPE}" -exec echo -e "bwa/mergedLibrary/macs/${PEAK_TYPE}/"{}"\\t0,0,178" \\; > ${prefix}_peaks.igv.txt - """ -} - -/* - * STEP 6.2 Annotate peaks with HOMER - */ -process MergedLibAnnotatePeaks { - tag "$name" - label "process_medium" - publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}", mode: 'copy' - - when: - params.macs_gsize - - input: - set val(name), file(peak) from ch_mlib_macs_homer - file fasta from ch_fasta - file gtf from ch_gtf - - output: - file "*.txt" into ch_mlib_macs_annotate - - script: - prefix = "${name}.mLb.clN" - """ - annotatePeaks.pl \\ - $peak \\ - $fasta \\ - -gid \\ - -gtf $gtf \\ - -cpu $task.cpus \\ - > ${prefix}_peaks.annotatePeaks.txt - """ -} - -/* - * STEP 6.3 Aggregated QC plots for peaks, FRiP and peak-to-gene annotation - */ -process MergedLibPeakQC { - label "process_medium" - publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/qc", mode: 'copy' - - when: - params.macs_gsize - - input: - file peaks from ch_mlib_macs_qc.collect{ it[1] } - file annos from ch_mlib_macs_annotate.collect() - file mlib_peak_annotation_header from ch_mlib_peak_annotation_header - - output: - file "*.{txt,pdf}" into ch_mlib_peak_qc - file "*.tsv" into ch_mlib_peak_qc_mqc - - script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ - suffix = 'mLb.clN' - """ - plot_macs_qc.r \\ - -i ${peaks.join(',')} \\ - -s ${peaks.join(',').replaceAll(".${suffix}_peaks.${PEAK_TYPE}","")} \\ - -o ./ \\ - -p macs_peak.${suffix} - - plot_homer_annotatepeaks.r \\ - -i ${annos.join(',')} \\ - -s ${annos.join(',').replaceAll(".${suffix}_peaks.annotatePeaks.txt","")} \\ - -o ./ \\ - -p macs_annotatePeaks.${suffix} - - cat $mlib_peak_annotation_header macs_annotatePeaks.${suffix}.summary.txt > macs_annotatePeaks.${suffix}.summary_mqc.tsv - """ -} - -/* - * STEP 6.4 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection - */ -process MergedLibConsensusPeakSet { - label 'process_long' - publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: 'copy', - saveAs: { filename -> - if (filename.endsWith(".igv.txt")) null - else filename - } - - when: - params.macs_gsize && (replicatesExist || multipleGroups) && !params.skip_consensus_peaks - - input: - file peaks from ch_mlib_macs_consensus.collect{ it[1] } - - output: - file "*.bed" into ch_mlib_macs_consensus_bed - file "*.saf" into ch_mlib_macs_consensus_saf - file "*.boolean.txt" into ch_mlib_macs_consensus_bool - file "*.intersect.{txt,plot.pdf}" into ch_mlib_macs_consensus_intersect - file "*igv.txt" into ch_mlib_macs_consensus_igv - - script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ - suffix = 'mLb.clN' - prefix = "consensus_peaks.${suffix}" - mergecols = params.narrow_peak ? (2..10).join(',') : (2..9).join(',') - collapsecols = params.narrow_peak ? (["collapse"]*9).join(',') : (["collapse"]*8).join(',') - expandparam = params.narrow_peak ? "--is_narrow_peak" : "" - """ - sort -T '.' -k1,1 -k2,2n ${peaks.collect{it.toString()}.sort().join(' ')} \\ - | mergeBed -c $mergecols -o $collapsecols > ${prefix}.txt - - macs2_merged_expand.py \\ - ${prefix}.txt \\ - ${peaks.collect{it.toString()}.sort().join(',').replaceAll("_peaks.${PEAK_TYPE}","")} \\ - ${prefix}.boolean.txt \\ - --min_replicates $params.min_reps_consensus \\ - $expandparam - - awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$1, \$2, \$3, \$4, "0", "+" }' ${prefix}.boolean.txt > ${prefix}.bed - - echo -e "GeneID\tChr\tStart\tEnd\tStrand" > ${prefix}.saf - awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$4, \$1, \$2, \$3, "+" }' ${prefix}.boolean.txt >> ${prefix}.saf - - sed -i 's/.${suffix}//g' ${prefix}.boolean.intersect.txt - plot_peak_intersect.r -i ${prefix}.boolean.intersect.txt -o ${prefix}.boolean.intersect.plot.pdf - - find * -type f -name "${prefix}.bed" -exec echo -e "bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus/"{}"\\t0,0,0" \\; > ${prefix}.bed.igv.txt - """ -} - -/* - * STEP 6.5 Annotate consensus peaks with HOMER, and add annotation to boolean output file - */ -process MergedLibConsensusPeakSetAnnotate { - label "process_medium" - publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: 'copy' - - when: - params.macs_gsize && (replicatesExist || multipleGroups) && !params.skip_consensus_peaks - - input: - file bed from ch_mlib_macs_consensus_bed - file bool from ch_mlib_macs_consensus_bool - file fasta from ch_fasta - file gtf from ch_gtf - - output: - file "*.annotatePeaks.txt" into ch_mlib_macs_consensus_annotate - - script: - prefix = "consensus_peaks.mLb.clN" - """ - annotatePeaks.pl \\ - $bed \\ - $fasta \\ - -gid \\ - -gtf $gtf \\ - -cpu $task.cpus \\ - > ${prefix}.annotatePeaks.txt - - cut -f2- ${prefix}.annotatePeaks.txt | awk 'NR==1; NR > 1 {print \$0 | "sort -T '.' -k1,1 -k2,2n"}' | cut -f6- > tmp.txt - paste $bool tmp.txt > ${prefix}.boolean.annotatePeaks.txt - """ -} - -/* - * STEP 6.6 Count reads in consensus peaks with featureCounts and perform differential analysis with DESeq2 - */ -process MergedLibConsensusPeakSetDESeq { - label 'process_medium' - publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus/deseq2", mode: 'copy', - saveAs: { filename -> - if (filename.endsWith(".igv.txt")) null - else filename - } - - when: - params.macs_gsize && replicatesExist && multipleGroups && !params.skip_consensus_peaks - - input: - file bams from ch_mlib_name_bam_mlib_counts.collect{ it[1] } - file saf from ch_mlib_macs_consensus_saf.collect() - file mlib_deseq2_pca_header from ch_mlib_deseq2_pca_header - file mlib_deseq2_clustering_header from ch_mlib_deseq2_clustering_header - - output: - file "*featureCounts.txt" into ch_mlib_macs_consensus_counts - file "*featureCounts.txt.summary" into ch_mlib_macs_consensus_counts_mqc - file "*.{RData,results.txt,pdf,log}" into ch_mlib_macs_consensus_deseq_results - file "sizeFactors" into ch_mlib_macs_consensus_deseq_factors - file "*vs*/*.{pdf,txt}" into ch_mlib_macs_consensus_deseq_comp_results - file "*vs*/*.bed" into ch_mlib_macs_consensus_deseq_comp_bed - file "*igv.txt" into ch_mlib_macs_consensus_deseq_comp_igv - file "*.tsv" into ch_mlib_macs_consensus_deseq_mqc - - script: - prefix = "consensus_peaks.mLb.clN" - bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() - bam_ext = params.single_end ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" - pe_params = params.single_end ? '' : "-p --donotsort" - """ - featureCounts \\ - -F SAF \\ - -O \\ - --fracOverlap 0.2 \\ - -T $task.cpus \\ - $pe_params \\ - -a $saf \\ - -o ${prefix}.featureCounts.txt \\ - ${bam_files.join(' ')} - - featurecounts_deseq2.r -i ${prefix}.featureCounts.txt -b '$bam_ext' -o ./ -p $prefix -s .mLb - - cat $mlib_deseq2_pca_header ${prefix}.pca.vals.txt > ${prefix}.pca.vals_mqc.tsv - cat $mlib_deseq2_clustering_header ${prefix}.sample.dists.txt > ${prefix}.sample.dists_mqc.tsv - - find * -type f -name "*.FDR0.05.results.bed" -exec echo -e "bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus/deseq2/"{}"\\t255,0,0" \\; > ${prefix}.igv.txt - """ -} - -/* - * STEP 6.7 Run ataqv on BAM file and corresponding peaks - */ -process MergedLibAtaqv { - tag "$name" - label 'process_medium' - publishDir "${params.outdir}/bwa/mergedLibrary/ataqv/${PEAK_TYPE}", mode: 'copy' - - when: - !params.skip_ataqv - - input: - set val(name), file(bam), file(peak) from ch_mlib_bam_ataqv.join(ch_mlib_macs_ataqv, by: [0]) - file autosomes from ch_genome_autosomes.collect() - file tss_bed from ch_tss_bed - - output: - file "*.json" into ch_mlib_ataqv - - script: - peak_param = params.macs_gsize ? "--peak-file ${peak}" : "" - mito_param = params.mito_name ? "--mitochondrial-reference-name ${params.mito_name}" : "" - """ - ataqv \\ - --threads $task.cpus \\ - $peak_param \\ - --tss-file $tss_bed \\ - --metrics-file ${name}.ataqv.json \\ - --name $name \\ - --ignore-read-groups \\ - --autosomal-reference-file $autosomes \\ - $mito_param \\ - NA \\ - ${bam[0]} - """ -} - -/* - * STEP 6.8 run ataqv mkarv on all JSON files to render web app - */ -process MergedLibAtaqvMkarv { - label 'process_medium' - publishDir "${params.outdir}/bwa/mergedLibrary/ataqv/${PEAK_TYPE}", mode: 'copy' - - when: - !params.skip_ataqv - - input: - file json from ch_mlib_ataqv.collect() - - output: - file "html" into ch_mlib_ataqv_mkarv - - script: - """ - mkarv \\ - --concurrency $task.cpus \\ - --force \\ - ./html/ \\ - ${json.join(' ')} - """ -} - -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// -/* -- -- */ -/* -- MERGE REPLICATE BAM -- */ -/* -- -- */ -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// - -/* - * STEP 7 Merge library BAM files across all replicates - */ -ch_mlib_rm_orphan_bam_mrep - .map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] } - .groupTuple(by: [0]) - .map { it -> [ it[0], it[1].flatten() ] } - .set { ch_mlib_rm_orphan_bam_mrep } - -process MergedRepBAM { - tag "$name" - label 'process_medium' - publishDir "${params.outdir}/bwa/mergedReplicate", mode: 'copy', - saveAs: { filename -> - if (filename.endsWith(".flagstat")) "samtools_stats/$filename" - else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" - else if (filename.endsWith(".stats")) "samtools_stats/$filename" - else if (filename.endsWith(".metrics.txt")) "picard_metrics/$filename" - else filename - } - - input: - set val(name), file(bams) from ch_mlib_rm_orphan_bam_mrep - - output: - set val(name), file("*${prefix}.sorted.{bam,bam.bai}") into ch_mrep_bam_bigwig, - ch_mrep_bam_macs - set val(name), file("*.flagstat") into ch_mrep_bam_flagstat_bigwig, - ch_mrep_bam_flagstat_macs, - ch_mrep_bam_flagstat_mqc - file "*.{idxstats,stats}" into ch_mrep_bam_stats_mqc - file "*.txt" into ch_mrep_bam_metrics_mqc - - when: - !params.skip_merge_replicates && replicatesExist - - script: - prefix = "${name}.mRp.clN" - bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() - def avail_mem = 3 - if (!task.memory) { - log.info "[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." - } else { - avail_mem = task.memory.toGiga() - } - if (bam_files.size() > 1) { - """ - picard -Xmx${avail_mem}g MergeSamFiles \\ - ${'INPUT='+bam_files.join(' INPUT=')} \\ - OUTPUT=${name}.sorted.bam \\ - SORT_ORDER=coordinate \\ - VALIDATION_STRINGENCY=LENIENT \\ - TMP_DIR=tmp - samtools index ${name}.sorted.bam - - picard -Xmx${avail_mem}g MarkDuplicates \\ - INPUT=${name}.sorted.bam \\ - OUTPUT=${prefix}.sorted.bam \\ - ASSUME_SORTED=true \\ - REMOVE_DUPLICATES=true \\ - METRICS_FILE=${prefix}.MarkDuplicates.metrics.txt \\ - VALIDATION_STRINGENCY=LENIENT \\ - TMP_DIR=tmp - - samtools index ${prefix}.sorted.bam - samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat - samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats - samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats - """ - } else { - """ - ln -s ${bams[0]} ${prefix}.sorted.bam - ln -s ${bams[1]} ${prefix}.sorted.bam.bai - touch ${prefix}.MarkDuplicates.metrics.txt - samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat - samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats - samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats - """ - } -} - -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// -/* -- -- */ -/* -- MERGE REPLICATE BAM POST-ANALYSIS -- */ -/* -- -- */ -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// - -/* - * STEP 8.1 Read depth normalised bigWig - */ -process MergedRepBigWig { - tag "$name" - label 'process_medium' - publishDir "${params.outdir}/bwa/mergedReplicate/bigwig", mode: 'copy', - saveAs: { filename -> - if (filename.endsWith("scale_factor.txt")) "scale/$filename" - else if (filename.endsWith(".bigWig")) "$filename" - else null - } - - when: - !params.skip_merge_replicates && replicatesExist - - input: - set val(name), file(bam), file(flagstat) from ch_mrep_bam_bigwig.join(ch_mrep_bam_flagstat_bigwig, by: [0]) - file sizes from ch_genome_sizes_mrep_bigwig.collect() - - output: - set val(name), file("*.bigWig") into ch_mrep_bigwig - file "*scale_factor.txt" into ch_mrep_bigwig_scale - file "*igv.txt" into ch_mrep_bigwig_igv - - script: - prefix = "${name}.mRp.clN" - pe_fragment = params.single_end ? "" : "-pc" - extend = (params.single_end && params.fragment_size > 0) ? "-fs ${params.fragment_size}" : '' - """ - SCALE_FACTOR=\$(grep 'mapped (' $flagstat | awk '{print 1000000/\$1}') - echo \$SCALE_FACTOR > ${prefix}.scale_factor.txt - genomeCoverageBed -ibam ${bam[0]} -bg -scale \$SCALE_FACTOR $pe_fragment $extend | sort -T '.' -k1,1 -k2,2n > ${prefix}.bedGraph - - bedGraphToBigWig ${prefix}.bedGraph $sizes ${prefix}.bigWig - - find * -type f -name "*.bigWig" -exec echo -e "bwa/mergedReplicate/bigwig/"{}"\\t0,0,178" \\; > ${prefix}.bigWig.igv.txt - """ -} - -/* - * STEP 8.2 Call peaks with MACS2 and calculate FRiP score - */ -process MergedRepMACSCallPeak { - tag "$name" - label 'process_medium' - publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}", mode: 'copy', - saveAs: { filename -> - if (filename.endsWith(".tsv")) "qc/$filename" - else if (filename.endsWith(".igv.txt")) null - else filename - } - - when: - !params.skip_merge_replicates && replicatesExist && params.macs_gsize - - input: - set val(name), file(bam), file(flagstat) from ch_mrep_bam_macs.join(ch_mrep_bam_flagstat_macs, by: [0]) - file mrep_peak_count_header from ch_mrep_peak_count_header - file mrep_frip_score_header from ch_mrep_frip_score_header - - output: - set val(name), file("*.{bed,xls,gappedPeak,bdg}") into ch_mrep_macs_output - set val(name), file("*$PEAK_TYPE") into ch_mrep_macs_homer, - ch_mrep_macs_qc, - ch_mrep_macs_consensus - file "*igv.txt" into ch_mrep_macs_igv - file "*_mqc.tsv" into ch_mrep_macs_mqc - - script: - prefix = "${name}.mRp.clN" - broad = params.narrow_peak ? '' : "--broad --broad-cutoff ${params.broad_cutoff}" - format = params.single_end ? "BAM" : "BAMPE" - pileup = params.save_macs_pileup ? "-B --SPMR" : "" - """ - macs2 callpeak \\ - -t ${bam[0]} \\ - $broad \\ - -f $format \\ - -g $params.macs_gsize \\ - -n $prefix \\ - $pileup \\ - --keep-dup all \\ - --nomodel - - cat ${prefix}_peaks.${PEAK_TYPE} | wc -l | awk -v OFS='\t' '{ print "${name}", \$1 }' | cat $mrep_peak_count_header - > ${prefix}_peaks.count_mqc.tsv - - READS_IN_PEAKS=\$(intersectBed -a ${bam[0]} -b ${prefix}_peaks.${PEAK_TYPE} -bed -c -f 0.20 | awk -F '\t' '{sum += \$NF} END {print sum}') - grep 'mapped (' $flagstat | awk -v a="\$READS_IN_PEAKS" -v OFS='\t' '{print "${name}", a/\$1}' | cat $mrep_frip_score_header - > ${prefix}_peaks.FRiP_mqc.tsv - - find * -type f -name "*.${PEAK_TYPE}" -exec echo -e "bwa/mergedReplicate/macs/${PEAK_TYPE}/"{}"\\t0,0,178" \\; > ${prefix}_peaks.igv.txt - """ -} - -/* - * STEP 8.3 Annotate peaks with HOMER - */ -process MergedRepAnnotatePeaks { - tag "$name" - label "process_medium" - publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}", mode: 'copy' - - when: - !params.skip_merge_replicates && replicatesExist && params.macs_gsize - - input: - set val(name), file(peak) from ch_mrep_macs_homer - file fasta from ch_fasta - file gtf from ch_gtf - - output: - file "*.txt" into ch_mrep_macs_annotate - - script: - prefix = "${name}.mRp.clN" - """ - annotatePeaks.pl \\ - $peak \\ - $fasta \\ - -gid \\ - -gtf $gtf \\ - -cpu $task.cpus \\ - > ${prefix}_peaks.annotatePeaks.txt - """ -} - -/* - * STEP 8.4 Aggregated QC plots for peaks, FRiP and peak-to-gene annotation - */ -process MergedRepPeakQC { - label "process_medium" - publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/qc", mode: 'copy' - - when: - !params.skip_merge_replicates && replicatesExist && params.macs_gsize - - input: - file peaks from ch_mrep_macs_qc.collect{ it[1] } - file annos from ch_mrep_macs_annotate.collect() - file mrep_peak_annotation_header from ch_mrep_peak_annotation_header - - output: - file "*.{txt,pdf}" into ch_mrep_peak_qc - file "*.tsv" into ch_mrep_peak_qc_mqc - - script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ - suffix = 'mRp.clN' - """ - plot_macs_qc.r \\ - -i ${peaks.join(',')} \\ - -s ${peaks.join(',').replaceAll(".${suffix}_peaks.${PEAK_TYPE}","")} \\ - -o ./ \\ - -p macs_peak.${suffix} - - plot_homer_annotatepeaks.r \\ - -i ${annos.join(',')} \\ - -s ${annos.join(',').replaceAll(".${suffix}_peaks.annotatePeaks.txt","")} \\ - -o ./ \\ - -p macs_annotatePeaks.${suffix} - - cat $mrep_peak_annotation_header macs_annotatePeaks.${suffix}.summary.txt > macs_annotatePeaks.${suffix}.summary_mqc.tsv - """ -} - -/* - * STEP 8.5 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection - */ -process MergedRepConsensusPeakSet { - label 'process_long' - publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: 'copy', - saveAs: { filename -> - if (filename.endsWith(".igv.txt")) null - else filename - } - - when: - !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks - - input: - file peaks from ch_mrep_macs_consensus.collect{ it[1] } - - output: - file "*.bed" into ch_mrep_macs_consensus_bed - file "*.saf" into ch_mrep_macs_consensus_saf - file "*.boolean.txt" into ch_mrep_macs_consensus_bool - file "*.intersect.{txt,plot.pdf}" into ch_mrep_macs_consensus_intersect - file "*igv.txt" into ch_mrep_macs_consensus_igv - - script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ - suffix = 'mRp.clN' - prefix = "consensus_peaks.${suffix}" - mergecols = params.narrow_peak ? (2..10).join(',') : (2..9).join(',') - collapsecols = params.narrow_peak ? (["collapse"]*9).join(',') : (["collapse"]*8).join(',') - expandparam = params.narrow_peak ? "--is_narrow_peak" : "" - """ - sort -T '.' -k1,1 -k2,2n ${peaks.collect{it.toString()}.sort().join(' ')} \\ - | mergeBed -c $mergecols -o $collapsecols > ${prefix}.txt - - macs2_merged_expand.py \\ - ${prefix}.txt \\ - ${peaks.collect{it.toString()}.sort().join(',').replaceAll("_peaks.${PEAK_TYPE}","")} \\ - ${prefix}.boolean.txt \\ - $expandparam - - awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$1, \$2, \$3, \$4, "0", "+" }' ${prefix}.boolean.txt > ${prefix}.bed - - echo -e "GeneID\tChr\tStart\tEnd\tStrand" > ${prefix}.saf - awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$4, \$1, \$2, \$3, "+" }' ${prefix}.boolean.txt >> ${prefix}.saf - - sed -i 's/.${suffix}//g' ${prefix}.boolean.intersect.txt - plot_peak_intersect.r -i ${prefix}.boolean.intersect.txt -o ${prefix}.boolean.intersect.plot.pdf - - find * -type f -name "${prefix}.bed" -exec echo -e "bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus/"{}"\\t0,0,0" \\; > ${prefix}.bed.igv.txt - """ -} - -/* - * STEP 8.6 Annotate consensus peaks with HOMER, and add annotation to boolean output file - */ -process MergedRepConsensusPeakSetAnnotate { - label "process_medium" - publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: 'copy' - - when: - !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks - - input: - file bed from ch_mrep_macs_consensus_bed - file bool from ch_mrep_macs_consensus_bool - file fasta from ch_fasta - file gtf from ch_gtf - - output: - file "*.annotatePeaks.txt" into ch_mrep_macs_consensus_annotate - - script: - prefix = "consensus_peaks.mRp.clN" - """ - annotatePeaks.pl \\ - $bed \\ - $fasta \\ - -gid \\ - -gtf $gtf \\ - -cpu $task.cpus \\ - > ${prefix}.annotatePeaks.txt - - cut -f2- ${prefix}.annotatePeaks.txt | awk 'NR==1; NR > 1 {print \$0 | "sort -T '.' -k1,1 -k2,2n"}' | cut -f6- > tmp.txt - paste $bool tmp.txt > ${prefix}.boolean.annotatePeaks.txt - """ -} - -/* - * STEP 8.7 Count reads in consensus peaks with featureCounts and perform differential analysis with DESeq2 - */ -process MergedRepConsensusPeakSetDESeq { - label 'process_medium' - publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus/deseq2", mode: 'copy', - saveAs: { filename -> - if (filename.endsWith(".igv.txt")) null - else filename - } - - when: - !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks - - input: - file bams from ch_mlib_name_bam_mrep_counts.collect{ it[1] } - file saf from ch_mrep_macs_consensus_saf.collect() - file mrep_deseq2_pca_header from ch_mrep_deseq2_pca_header - file mrep_deseq2_clustering_header from ch_mrep_deseq2_clustering_header - - output: - file "*featureCounts.txt" into ch_mrep_macs_consensus_counts - file "*featureCounts.txt.summary" into ch_mrep_macs_consensus_counts_mqc - file "*.{RData,results.txt,pdf,log}" into ch_mrep_macs_consensus_deseq_results - file "sizeFactors" into ch_mrep_macs_consensus_deseq_factors - file "*vs*/*.{pdf,txt}" into ch_mrep_macs_consensus_deseq_comp_results - file "*vs*/*.bed" into ch_mrep_macs_consensus_deseq_comp_bed - file "*igv.txt" into ch_mrep_macs_consensus_deseq_comp_igv - file "*.tsv" into ch_mrep_macs_consensus_deseq_mqc - - script: - prefix = "consensus_peaks.mRp.clN" - bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() - bam_ext = params.single_end ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" - pe_params = params.single_end ? '' : "-p --donotsort" - """ - featureCounts \\ - -F SAF \\ - -O \\ - --fracOverlap 0.2 \\ - -T $task.cpus \\ - $pe_params \\ - -a $saf \\ - -o ${prefix}.featureCounts.txt \\ - ${bam_files.join(' ')} - - featurecounts_deseq2.r -i ${prefix}.featureCounts.txt -b '$bam_ext' -o ./ -p $prefix -s .mLb - - cat $mrep_deseq2_pca_header ${prefix}.pca.vals.txt > ${prefix}.pca.vals_mqc.tsv - cat $mrep_deseq2_clustering_header ${prefix}.sample.dists.txt > ${prefix}.sample.dists_mqc.tsv - - find * -type f -name "*.FDR0.05.results.bed" -exec echo -e "bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus/deseq2/"{}"\\t255,0,0" \\; > ${prefix}.igv.txt - """ -} - -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// -/* -- -- */ -/* -- IGV -- */ -/* -- -- */ -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// - -/* - * STEP 9 - Create IGV session file - */ -process IGV { - publishDir "${params.outdir}/igv/${PEAK_TYPE}", mode: 'copy' - - when: - !params.skip_igv - - input: - file fasta from ch_fasta - - file bigwigs from ch_mlib_bigwig_igv.collect().ifEmpty([]) - file peaks from ch_mlib_macs_igv.collect().ifEmpty([]) - file consensus_peaks from ch_mlib_macs_consensus_igv.collect().ifEmpty([]) - file differential_peaks from ch_mlib_macs_consensus_deseq_comp_igv.collect().ifEmpty([]) - - file rbigwigs from ch_mrep_bigwig_igv.collect().ifEmpty([]) - file rpeaks from ch_mrep_macs_igv.collect().ifEmpty([]) - file rconsensus_peaks from ch_mrep_macs_consensus_igv.collect().ifEmpty([]) - file rdifferential_peaks from ch_mrep_macs_consensus_deseq_comp_igv.collect().ifEmpty([]) - - output: - file "*.{txt,xml}" into ch_igv_session - - script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ - """ - cat *.txt > igv_files.txt - igv_files_to_session.py igv_session.xml igv_files.txt ../../reference_genome/${fasta.getName()} --path_prefix '../../' - """ -} - -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// -/* -- -- */ -/* -- MULTIQC -- */ -/* -- -- */ -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// - -/* - * Parse software version numbers - */ -process get_software_versions { - publishDir "${params.outdir}/pipeline_info", mode: 'copy', - saveAs: { filename -> - if (filename.indexOf(".csv") > 0) filename - else null - } - - output: - file 'software_versions_mqc.yaml' into ch_software_versions_mqc - file "software_versions.csv" - - script: - """ - echo $workflow.manifest.version > v_pipeline.txt - echo $workflow.nextflow.version > v_nextflow.txt - fastqc --version > v_fastqc.txt - trim_galore --version > v_trim_galore.txt - echo \$(bwa 2>&1) > v_bwa.txt - samtools --version > v_samtools.txt - bedtools --version > v_bedtools.txt - echo \$(bamtools --version 2>&1) > v_bamtools.txt - echo \$(plotFingerprint --version 2>&1) > v_deeptools.txt || true - picard MarkDuplicates --version &> v_picard.txt || true - echo \$(R --version 2>&1) > v_R.txt - python -c "import pysam; print(pysam.__version__)" > v_pysam.txt - echo \$(macs2 --version 2>&1) > v_macs2.txt - touch v_homer.txt - echo \$(ataqv --version 2>&1) > v_ataqv.txt - echo \$(featureCounts -v 2>&1) > v_featurecounts.txt - preseq &> v_preseq.txt - multiqc --version > v_multiqc.txt - scrape_software_versions.py &> software_versions_mqc.yaml - """ -} - -def create_workflow_summary(summary) { - - def yaml_file = workDir.resolve('workflow_summary_mqc.yaml') - yaml_file.text = """ - id: 'nf-core-atacseq-summary' - description: " - this information is collected when the pipeline is started." - section_name: 'nf-core/atacseq Workflow Summary' - section_href: 'https://github.com/nf-core/atacseq' - plot_type: 'html' - data: | -
-${summary.collect { k,v -> "
$k
${v ?: 'N/A'}
" }.join("\n")} -
- """.stripIndent() - - return yaml_file -} - -/* - * STEP 10 - MultiQC - */ -process MultiQC { - publishDir "${params.outdir}/multiqc/${PEAK_TYPE}", mode: 'copy' - - when: - !params.skip_multiqc - - input: - file multiqc_config from ch_multiqc_config - file readme from ch_anno_readme.collect().ifEmpty([]) - - file ('software_versions/*') from ch_software_versions_mqc.collect() - file ('workflow_summary/*') from create_workflow_summary(summary) - - file ('fastqc/*') from ch_fastqc_reports_mqc.collect().ifEmpty([]) - file ('trimgalore/*') from ch_trimgalore_results_mqc.collect().ifEmpty([]) - file ('trimgalore/fastqc/*') from ch_trimgalore_fastqc_reports_mqc.collect().ifEmpty([]) - - file ('alignment/library/*') from ch_sort_bam_flagstat_mqc.collect() - - file ('alignment/mergedLibrary/*') from ch_mlib_bam_stats_mqc.collect() - file ('alignment/mergedLibrary/*') from ch_mlib_rm_orphan_flagstat_mqc.collect{it[1]} - file ('alignment/mergedLibrary/*') from ch_mlib_rm_orphan_stats_mqc.collect() - file ('alignment/mergedLibrary/picard_metrics/*') from ch_mlib_bam_metrics_mqc.collect() - file ('alignment/mergedLibrary/picard_metrics/*') from ch_mlib_collectmetrics_mqc.collect() - file ('macs/mergedLibrary/*') from ch_mlib_macs_mqc.collect().ifEmpty([]) - file ('macs/mergedLibrary/*') from ch_mlib_peak_qc_mqc.collect().ifEmpty([]) - file ('macs/mergedLibrary/consensus/*') from ch_mlib_macs_consensus_counts_mqc.collect().ifEmpty([]) - file ('macs/mergedLibrary/consensus/*') from ch_mlib_macs_consensus_deseq_mqc.collect().ifEmpty([]) - file ('preseq/*') from ch_mlib_preseq_mqc.collect().ifEmpty([]) - file ('deeptools/*') from ch_mlib_plotprofile_mqc.collect().ifEmpty([]) - file ('deeptools/*') from ch_mlib_plotfingerprint_mqc.collect().ifEmpty([]) - - file ('alignment/mergedReplicate/*') from ch_mrep_bam_flagstat_mqc.collect{it[1]}.ifEmpty([]) - file ('alignment/mergedReplicate/*') from ch_mrep_bam_stats_mqc.collect().ifEmpty([]) - file ('alignment/mergedReplicate/*') from ch_mrep_bam_metrics_mqc.collect().ifEmpty([]) - file ('macs/mergedReplicate/*') from ch_mrep_macs_mqc.collect().ifEmpty([]) - file ('macs/mergedReplicate/*') from ch_mrep_peak_qc_mqc.collect().ifEmpty([]) - file ('macs/mergedReplicate/consensus/*') from ch_mrep_macs_consensus_counts_mqc.collect().ifEmpty([]) - file ('macs/mergedReplicate/consensus/*') from ch_mrep_macs_consensus_deseq_mqc.collect().ifEmpty([]) - - output: - file "*multiqc_report.html" into ch_multiqc_report - file "*_data" - file "multiqc_plots" - - script: - rtitle = custom_runName ? "--title \"$custom_runName\"" : '' - rfilename = custom_runName ? "--filename " + custom_runName.replaceAll('\\W','_').replaceAll('_+','_') + "_multiqc_report" : '' - """ - multiqc . -f $rtitle $rfilename --config $multiqc_config \\ - -m custom_content -m fastqc -m cutadapt -m samtools -m picard -m preseq -m featureCounts -m deeptools - """ -} - -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// -/* -- -- */ -/* -- REPORTS/DOCUMENTATION -- */ -/* -- -- */ -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// - -/* - * STEP 11 - Output description HTML - */ -process output_documentation { - publishDir "${params.outdir}/Documentation", mode: 'copy' - - input: - file output_docs from ch_output_docs - - output: - file "results_description.html" - - script: - """ - markdown_to_html.r $output_docs results_description.html - """ -} - -/* - * Completion e-mail notification - */ -workflow.onComplete { - - // Set up the e-mail variables - def subject = "[nf-core/atacseq] Successful: $workflow.runName" - if (!workflow.success) { - subject = "[nf-core/atacseq] FAILED: $workflow.runName" - } - def email_fields = [:] - email_fields['version'] = workflow.manifest.version - email_fields['runName'] = custom_runName ?: workflow.runName - email_fields['success'] = workflow.success - email_fields['dateComplete'] = workflow.complete - email_fields['duration'] = workflow.duration - email_fields['exitStatus'] = workflow.exitStatus - email_fields['errorMessage'] = (workflow.errorMessage ?: 'None') - email_fields['errorReport'] = (workflow.errorReport ?: 'None') - email_fields['commandLine'] = workflow.commandLine - email_fields['projectDir'] = workflow.projectDir - email_fields['summary'] = summary - email_fields['summary']['Date Started'] = workflow.start - email_fields['summary']['Date Completed'] = workflow.complete - email_fields['summary']['Pipeline script file path'] = workflow.scriptFile - email_fields['summary']['Pipeline script hash ID'] = workflow.scriptId - if (workflow.repository) email_fields['summary']['Pipeline repository Git URL'] = workflow.repository - if (workflow.commitId) email_fields['summary']['Pipeline repository Git Commit'] = workflow.commitId - if (workflow.revision) email_fields['summary']['Pipeline Git branch/tag'] = workflow.revision - if (workflow.container) email_fields['summary']['Docker image'] = workflow.container - email_fields['summary']['Nextflow Version'] = workflow.nextflow.version - email_fields['summary']['Nextflow Build'] = workflow.nextflow.build - email_fields['summary']['Nextflow Compile Timestamp'] = workflow.nextflow.timestamp - - // On success try attach the multiqc report - def mqc_report = null - try { - if (workflow.success) { - mqc_report = ch_multiqc_report.getVal() - if (mqc_report.getClass() == ArrayList) { - log.warn "[nf-core/atacseq] Found multiple reports from process 'multiqc', will use only one" - mqc_report = mqc_report[0] - } - } - } catch (all) { - log.warn "[nf-core/atacseq] Could not attach MultiQC report to summary email" - } - - // Check if we are only sending emails on failure - email_address = params.email - if (!params.email && params.email_on_fail && !workflow.success) { - email_address = params.email_on_fail - } - - // Render the TXT template - def engine = new groovy.text.GStringTemplateEngine() - def tf = new File("$baseDir/assets/email_template.txt") - def txt_template = engine.createTemplate(tf).make(email_fields) - def email_txt = txt_template.toString() - - // Render the HTML template - def hf = new File("$baseDir/assets/email_template.html") - def html_template = engine.createTemplate(hf).make(email_fields) - def email_html = html_template.toString() - - // Render the sendmail template - def smail_fields = [ email: email_address, subject: subject, email_txt: email_txt, email_html: email_html, baseDir: "$baseDir", mqcFile: mqc_report, mqcMaxSize: params.max_multiqc_email_size.toBytes() ] - def sf = new File("$baseDir/assets/sendmail_template.txt") - def sendmail_template = engine.createTemplate(sf).make(smail_fields) - def sendmail_html = sendmail_template.toString() - - // Send the HTML e-mail - if (email_address) { - try { - if (params.plaintext_email) { throw GroovyException('Send plaintext e-mail, not HTML') } - // Try to send HTML e-mail using sendmail - [ 'sendmail', '-t' ].execute() << sendmail_html - log.info "[nf-core/atacseq] Sent summary e-mail to $email_address (sendmail)" - } catch (all) { - // Catch failures and try with plaintext - [ 'mail', '-s', subject, email_address ].execute() << email_txt - log.info "[nf-core/atacseq] Sent summary e-mail to $email_address (mail)" - } - } - - // Write summary e-mail HTML to a file - def output_d = new File("${params.outdir}/pipeline_info/") - if (!output_d.exists()) { - output_d.mkdirs() - } - def output_hf = new File(output_d, "pipeline_report.html") - output_hf.withWriter { w -> w << email_html } - def output_tf = new File(output_d, "pipeline_report.txt") - output_tf.withWriter { w -> w << email_txt } - - c_reset = params.monochrome_logs ? '' : "\033[0m"; - c_purple = params.monochrome_logs ? '' : "\033[0;35m"; - c_green = params.monochrome_logs ? '' : "\033[0;32m"; - c_red = params.monochrome_logs ? '' : "\033[0;31m"; - - if (workflow.stats.ignoredCount > 0 && workflow.success) { - log.info "${c_purple}Warning, pipeline completed, but with errored process(es) ${c_reset}" - log.info "${c_red}Number of ignored errored process(es) : ${workflow.stats.ignoredCount} ${c_reset}" - log.info "${c_green}Number of successfully ran process(es) : ${workflow.stats.succeedCount} ${c_reset}" - } - - if (workflow.success) { - log.info "${c_purple}[nf-core/atacseq]${c_green} Pipeline completed successfully${c_reset}" - } else { - checkHostname() - log.info "${c_purple}[nf-core/atacseq]${c_red} Pipeline completed with errors${c_reset}" - } - -} +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// /* -- -- */ +// /* -- FASTQ QC -- */ +// /* -- -- */ +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// +// /* +// * STEP 1 - FastQC +// */ +// process FastQC { +// tag "$name" +// label 'process_medium' +// publishDir "${params.outdir}/fastqc", mode: 'copy', +// saveAs: { filename -> +// filename.endsWith(".zip") ? "zips/$filename" : "$filename" +// } +// +// when: +// !params.skip_fastqc +// +// input: +// set val(name), file(reads) from ch_raw_reads_fastqc +// +// output: +// file "*.{zip,html}" into ch_fastqc_reports_mqc +// +// script: +// // Added soft-links to original fastqs for consistent naming in MultiQC +// if (params.single_end) { +// """ +// [ ! -f ${name}.fastq.gz ] && ln -s $reads ${name}.fastq.gz +// fastqc -q -t $task.cpus ${name}.fastq.gz +// """ +// } else { +// """ +// [ ! -f ${name}_1.fastq.gz ] && ln -s ${reads[0]} ${name}_1.fastq.gz +// [ ! -f ${name}_2.fastq.gz ] && ln -s ${reads[1]} ${name}_2.fastq.gz +// fastqc -q -t $task.cpus ${name}_1.fastq.gz +// fastqc -q -t $task.cpus ${name}_2.fastq.gz +// """ +// } +// } +// +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// /* -- -- */ +// /* -- ADAPTER TRIMMING -- */ +// /* -- -- */ +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// +// /* +// * STEP 2 - Trim Galore! +// */ +// if (params.skip_trimming) { +// ch_trimmed_reads = ch_raw_reads_trimgalore +// ch_trimgalore_results_mqc = Channel.empty() +// ch_trimgalore_fastqc_reports_mqc = Channel.empty() +// } else { +// process TrimGalore { +// tag "$name" +// label 'process_high' +// publishDir "${params.outdir}/trim_galore", mode: 'copy', +// saveAs: { filename -> +// if (filename.endsWith(".html")) "fastqc/$filename" +// else if (filename.endsWith(".zip")) "fastqc/zips/$filename" +// else if (filename.endsWith("trimming_report.txt")) "logs/$filename" +// else params.save_trimmed ? filename : null +// } +// +// input: +// set val(name), file(reads) from ch_raw_reads_trimgalore +// +// output: +// set val(name), file("*.fq.gz") into ch_trimmed_reads +// file "*.txt" into ch_trimgalore_results_mqc +// file "*.{zip,html}" into ch_trimgalore_fastqc_reports_mqc +// +// script: +// // Calculate number of --cores for TrimGalore based on value of task.cpus +// // See: https://github.com/FelixKrueger/TrimGalore/blob/master/Changelog.md#version-060-release-on-1-mar-2019 +// // See: https://github.com/nf-core/atacseq/pull/65 +// def cores = 1 +// if (task.cpus) { +// cores = (task.cpus as int) - 4 +// if (params.single_end) cores = (task.cpus as int) - 3 +// if (cores < 1) cores = 1 +// if (cores > 4) cores = 4 +// } +// +// // Added soft-links to original fastqs for consistent naming in MultiQC +// c_r1 = params.clip_r1 > 0 ? "--clip_r1 ${params.clip_r1}" : '' +// c_r2 = params.clip_r2 > 0 ? "--clip_r2 ${params.clip_r2}" : '' +// tpc_r1 = params.three_prime_clip_r1 > 0 ? "--three_prime_clip_r1 ${params.three_prime_clip_r1}" : '' +// tpc_r2 = params.three_prime_clip_r2 > 0 ? "--three_prime_clip_r2 ${params.three_prime_clip_r2}" : '' +// nextseq = params.trim_nextseq > 0 ? "--nextseq ${params.trim_nextseq}" : '' +// +// // Added soft-links to original fastqs for consistent naming in MultiQC +// if (params.single_end) { +// """ +// [ ! -f ${name}.fastq.gz ] && ln -s $reads ${name}.fastq.gz +// trim_galore --cores $cores --fastqc --gzip $c_r1 $tpc_r1 $nextseq ${name}.fastq.gz +// """ +// } else { +// """ +// [ ! -f ${name}_1.fastq.gz ] && ln -s ${reads[0]} ${name}_1.fastq.gz +// [ ! -f ${name}_2.fastq.gz ] && ln -s ${reads[1]} ${name}_2.fastq.gz +// trim_galore --cores $cores --paired --fastqc --gzip $c_r1 $c_r2 $tpc_r1 $tpc_r2 $nextseq ${name}_1.fastq.gz ${name}_2.fastq.gz +// """ +// } +// } +// } +// +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// /* -- -- */ +// /* -- ALIGN -- */ +// /* -- -- */ +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// +// /* +// * STEP 3.1 - Align read 1 with bwa +// */ +// process BWAMem { +// tag "$name" +// label 'process_high' +// +// input: +// set val(name), file(reads) from ch_trimmed_reads +// file index from ch_bwa_index.collect() +// +// output: +// set val(name), file("*.bam") into ch_bwa_bam +// +// script: +// prefix = "${name}.Lb" +// rg = "\'@RG\\tID:${name}\\tSM:${name.split('_')[0..-2].join('_')}\\tPL:ILLUMINA\\tLB:${name}\\tPU:1\'" +// if (params.seq_center) { +// rg = "\'@RG\\tID:${name}\\tSM:${name.split('_')[0..-2].join('_')}\\tPL:ILLUMINA\\tLB:${name}\\tPU:1\\tCN:${params.seq_center}\'" +// } +// """ +// bwa mem \\ +// -t $task.cpus \\ +// -M \\ +// -R $rg \\ +// ${index}/${bwa_base} \\ +// $reads \\ +// | samtools view -@ $task.cpus -b -h -F 0x0100 -O BAM -o ${prefix}.bam - +// """ +// } +// +// /* +// * STEP 3.2 - Convert .bam to coordinate sorted .bam +// */ +// process SortBAM { +// tag "$name" +// label 'process_medium' +// if (params.save_align_intermeds) { +// publishDir path: "${params.outdir}/bwa/library", mode: 'copy', +// saveAs: { filename -> +// if (filename.endsWith(".flagstat")) "samtools_stats/$filename" +// else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" +// else if (filename.endsWith(".stats")) "samtools_stats/$filename" +// else filename +// } +// } +// +// input: +// set val(name), file(bam) from ch_bwa_bam +// +// output: +// set val(name), file("*.sorted.{bam,bam.bai}") into ch_sort_bam_merge +// file "*.{flagstat,idxstats,stats}" into ch_sort_bam_flagstat_mqc +// +// script: +// prefix = "${name}.Lb" +// """ +// samtools sort -@ $task.cpus -o ${prefix}.sorted.bam -T $name $bam +// samtools index ${prefix}.sorted.bam +// samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat +// samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats +// samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats +// """ +// } +// +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// /* -- -- */ +// /* -- MERGE LIBRARY BAM -- */ +// /* -- -- */ +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// +// /* +// * STEP 4.1 Merge BAM files for all libraries from same replicate +// */ +// ch_sort_bam_merge +// .map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] } +// .groupTuple(by: [0]) +// .map { it -> [ it[0], it[1].flatten() ] } +// .set { ch_sort_bam_merge } +// +// process MergedLibBAM { +// tag "$name" +// label 'process_medium' +// publishDir "${params.outdir}/bwa/mergedLibrary", mode: 'copy', +// saveAs: { filename -> +// if (filename.endsWith(".flagstat")) "samtools_stats/$filename" +// else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" +// else if (filename.endsWith(".stats")) "samtools_stats/$filename" +// else if (filename.endsWith(".metrics.txt")) "picard_metrics/$filename" +// else params.save_align_intermeds ? filename : null +// } +// +// input: +// set val(name), file(bams) from ch_sort_bam_merge +// +// output: +// set val(name), file("*${prefix}.sorted.{bam,bam.bai}") into ch_mlib_bam_filter, +// ch_mlib_bam_preseq, +// ch_mlib_bam_ataqv +// file "*.{flagstat,idxstats,stats}" into ch_mlib_bam_stats_mqc +// file "*.txt" into ch_mlib_bam_metrics_mqc +// +// script: +// prefix = "${name}.mLb.mkD" +// bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() +// def avail_mem = 3 +// if (!task.memory) { +// log.info "[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." +// } else { +// avail_mem = task.memory.toGiga() +// } +// if (bam_files.size() > 1) { +// """ +// picard -Xmx${avail_mem}g MergeSamFiles \\ +// ${'INPUT='+bam_files.join(' INPUT=')} \\ +// OUTPUT=${name}.sorted.bam \\ +// SORT_ORDER=coordinate \\ +// VALIDATION_STRINGENCY=LENIENT \\ +// TMP_DIR=tmp +// samtools index ${name}.sorted.bam +// +// picard -Xmx${avail_mem}g MarkDuplicates \\ +// INPUT=${name}.sorted.bam \\ +// OUTPUT=${prefix}.sorted.bam \\ +// ASSUME_SORTED=true \\ +// REMOVE_DUPLICATES=false \\ +// METRICS_FILE=${prefix}.MarkDuplicates.metrics.txt \\ +// VALIDATION_STRINGENCY=LENIENT \\ +// TMP_DIR=tmp +// +// samtools index ${prefix}.sorted.bam +// samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats +// samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat +// samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats +// """ +// } else { +// """ +// picard -Xmx${avail_mem}g MarkDuplicates \\ +// INPUT=${bam_files[0]} \\ +// OUTPUT=${prefix}.sorted.bam \\ +// ASSUME_SORTED=true \\ +// REMOVE_DUPLICATES=false \\ +// METRICS_FILE=${prefix}.MarkDuplicates.metrics.txt \\ +// VALIDATION_STRINGENCY=LENIENT \\ +// TMP_DIR=tmp +// +// samtools index ${prefix}.sorted.bam +// samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats +// samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat +// samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats +// """ +// } +// } +// +// /* +// * STEP 4.2 Filter BAM file at merged library-level +// */ +// process MergedLibBAMFilter { +// tag "$name" +// label 'process_medium' +// publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', +// saveAs: { filename -> +// if (params.single_end || params.save_align_intermeds) { +// if (filename.endsWith(".flagstat")) "samtools_stats/$filename" +// else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" +// else if (filename.endsWith(".stats")) "samtools_stats/$filename" +// else if (filename.endsWith(".sorted.bam")) filename +// else if (filename.endsWith(".sorted.bam.bai")) filename +// else null +// } +// } +// +// input: +// set val(name), file(bam) from ch_mlib_bam_filter +// file bed from ch_genome_filter_regions.collect() +// file bamtools_filter_config from ch_bamtools_filter_config +// +// output: +// set val(name), file("*.{bam,bam.bai}") into ch_mlib_filter_bam +// set val(name), file("*.flagstat") into ch_mlib_filter_bam_flagstat +// file "*.{idxstats,stats}" into ch_mlib_filter_bam_stats_mqc +// +// script: +// prefix = params.single_end ? "${name}.mLb.clN" : "${name}.mLb.flT" +// filter_params = params.single_end ? "-F 0x004" : "-F 0x004 -F 0x0008 -f 0x001" +// dup_params = params.keep_dups ? "" : "-F 0x0400" +// multimap_params = params.keep_multi_map ? "" : "-q 1" +// blacklist_params = params.blacklist ? "-L $bed" : "" +// name_sort_bam = params.single_end ? "" : "samtools sort -n -@ $task.cpus -o ${prefix}.bam -T $prefix ${prefix}.sorted.bam" +// """ +// samtools view \\ +// $filter_params \\ +// $dup_params \\ +// $multimap_params \\ +// $blacklist_params \\ +// -b ${bam[0]} \\ +// | bamtools filter \\ +// -out ${prefix}.sorted.bam \\ +// -script $bamtools_filter_config +// +// samtools index ${prefix}.sorted.bam +// samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat +// samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats +// samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats +// +// $name_sort_bam +// """ +// } +// +// /* +// * STEP 4.3 Remove orphan reads from paired-end BAM file +// */ +// if (params.single_end) { +// ch_mlib_filter_bam +// .into { ch_mlib_rm_orphan_bam_metrics; +// ch_mlib_rm_orphan_bam_bigwig; +// ch_mlib_rm_orphan_bam_macs; +// ch_mlib_rm_orphan_bam_plotfingerprint; +// ch_mlib_rm_orphan_bam_mrep; +// ch_mlib_name_bam_mlib_counts; +// ch_mlib_name_bam_mrep_counts } +// +// ch_mlib_filter_bam_flagstat +// .into { ch_mlib_rm_orphan_flagstat_bigwig; +// ch_mlib_rm_orphan_flagstat_macs; +// ch_mlib_rm_orphan_flagstat_mqc } +// +// ch_mlib_filter_bam_stats_mqc +// .set { ch_mlib_rm_orphan_stats_mqc } +// } else { +// process MergedLibBAMRemoveOrphan { +// tag "$name" +// label 'process_medium' +// publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', +// saveAs: { filename -> +// if (filename.endsWith(".flagstat")) "samtools_stats/$filename" +// else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" +// else if (filename.endsWith(".stats")) "samtools_stats/$filename" +// else if (filename.endsWith(".sorted.bam")) filename +// else if (filename.endsWith(".sorted.bam.bai")) filename +// else null +// } +// +// input: +// set val(name), file(bam) from ch_mlib_filter_bam +// +// output: +// set val(name), file("*.sorted.{bam,bam.bai}") into ch_mlib_rm_orphan_bam_metrics, +// ch_mlib_rm_orphan_bam_bigwig, +// ch_mlib_rm_orphan_bam_macs, +// ch_mlib_rm_orphan_bam_plotfingerprint, +// ch_mlib_rm_orphan_bam_mrep +// set val(name), file("${prefix}.bam") into ch_mlib_name_bam_mlib_counts, +// ch_mlib_name_bam_mrep_counts +// set val(name), file("*.flagstat") into ch_mlib_rm_orphan_flagstat_bigwig, +// ch_mlib_rm_orphan_flagstat_macs, +// ch_mlib_rm_orphan_flagstat_mqc +// file "*.{idxstats,stats}" into ch_mlib_rm_orphan_stats_mqc +// +// script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ +// prefix = "${name}.mLb.clN" +// """ +// bampe_rm_orphan.py ${bam[0]} ${prefix}.bam --only_fr_pairs +// +// samtools sort -@ $task.cpus -o ${prefix}.sorted.bam -T $prefix ${prefix}.bam +// samtools index ${prefix}.sorted.bam +// samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat +// samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats +// samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats +// """ +// } +// } +// +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// /* -- -- */ +// /* -- MERGE LIBRARY BAM POST-ANALYSIS -- */ +// /* -- -- */ +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// +// /* +// * STEP 5.1 preseq analysis after merging libraries and before filtering +// */ +// process MergedLibPreseq { +// tag "$name" +// label 'process_low' +// publishDir "${params.outdir}/bwa/mergedLibrary/preseq", mode: 'copy' +// +// when: +// !params.skip_preseq +// +// input: +// set val(name), file(bam) from ch_mlib_bam_preseq +// +// output: +// file "*.ccurve.txt" into ch_mlib_preseq_mqc +// +// script: +// prefix = "${name}.mLb.mkD" +// """ +// preseq lc_extrap -v -output ${prefix}.ccurve.txt -bam ${bam[0]} +// """ +// } +// +// /* +// * STEP 5.2 Picard CollectMultipleMetrics after merging libraries and filtering +// */ +// process MergedLibMetrics { +// tag "$name" +// label 'process_medium' +// publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', +// saveAs: { filename -> +// if (filename.endsWith("_metrics")) "picard_metrics/$filename" +// else if (filename.endsWith(".pdf")) "picard_metrics/pdf/$filename" +// else null +// } +// +// when: +// !params.skip_picard_metrics +// +// input: +// set val(name), file(bam) from ch_mlib_rm_orphan_bam_metrics +// file fasta from ch_fasta +// +// output: +// file "*_metrics" into ch_mlib_collectmetrics_mqc +// file "*.pdf" into ch_mlib_collectmetrics_pdf +// +// script: +// prefix = "${name}.mLb.clN" +// def avail_mem = 3 +// if (!task.memory) { +// log.info "[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." +// } else { +// avail_mem = task.memory.toGiga() +// } +// """ +// picard -Xmx${avail_mem}g CollectMultipleMetrics \\ +// INPUT=${bam[0]} \\ +// OUTPUT=${prefix}.CollectMultipleMetrics \\ +// REFERENCE_SEQUENCE=$fasta \\ +// VALIDATION_STRINGENCY=LENIENT \\ +// TMP_DIR=tmp +// """ +// } +// +// /* +// * STEP 5.3 Read depth normalised bigWig +// */ +// process MergedLibBigWig { +// tag "$name" +// label 'process_medium' +// publishDir "${params.outdir}/bwa/mergedLibrary/bigwig", mode: 'copy', +// saveAs: { filename -> +// if (filename.endsWith("scale_factor.txt")) "scale/$filename" +// else if (filename.endsWith(".bigWig")) "$filename" +// else null +// } +// +// input: +// set val(name), file(bam), file(flagstat) from ch_mlib_rm_orphan_bam_bigwig.join(ch_mlib_rm_orphan_flagstat_bigwig, by: [0]) +// file sizes from ch_genome_sizes_mlib_bigwig.collect() +// +// output: +// set val(name), file("*.bigWig") into ch_mlib_bigwig_plotprofile +// file "*scale_factor.txt" into ch_mlib_bigwig_scale +// file "*igv.txt" into ch_mlib_bigwig_igv +// +// script: +// prefix = "${name}.mLb.clN" +// pe_fragment = params.single_end ? "" : "-pc" +// extend = (params.single_end && params.fragment_size > 0) ? "-fs ${params.fragment_size}" : '' +// """ +// SCALE_FACTOR=\$(grep 'mapped (' $flagstat | awk '{print 1000000/\$1}') +// echo \$SCALE_FACTOR > ${prefix}.scale_factor.txt +// genomeCoverageBed -ibam ${bam[0]} -bg -scale \$SCALE_FACTOR $pe_fragment $extend | sort -T '.' -k1,1 -k2,2n > ${prefix}.bedGraph +// +// bedGraphToBigWig ${prefix}.bedGraph $sizes ${prefix}.bigWig +// +// find * -type f -name "*.bigWig" -exec echo -e "bwa/mergedLibrary/bigwig/"{}"\\t0,0,178" \\; > ${prefix}.bigWig.igv.txt +// """ +// } +// +// /* +// * STEP 5.4 generate gene body coverage plot with deepTools +// */ +// process MergedLibPlotProfile { +// tag "$name" +// label 'process_high' +// publishDir "${params.outdir}/bwa/mergedLibrary/deepTools/plotProfile", mode: 'copy' +// +// when: +// !params.skip_plot_profile +// +// input: +// set val(name), file(bigwig) from ch_mlib_bigwig_plotprofile +// file bed from ch_gene_bed +// +// output: +// file '*.{gz,pdf}' into ch_mlib_plotprofile_results +// file '*.plotProfile.tab' into ch_mlib_plotprofile_mqc +// +// script: +// prefix = "${name}.mLb.clN" +// """ +// computeMatrix scale-regions \\ +// --regionsFileName $bed \\ +// --scoreFileName $bigwig \\ +// --outFileName ${prefix}.computeMatrix.mat.gz \\ +// --outFileNameMatrix ${prefix}.computeMatrix.vals.mat.gz \\ +// --regionBodyLength 1000 \\ +// --beforeRegionStartLength 3000 \\ +// --afterRegionStartLength 3000 \\ +// --skipZeros \\ +// --smartLabels \\ +// --numberOfProcessors $task.cpus +// +// plotProfile --matrixFile ${prefix}.computeMatrix.mat.gz \\ +// --outFileName ${prefix}.plotProfile.pdf \\ +// --outFileNameData ${prefix}.plotProfile.tab +// """ +// } +// +// /* +// * STEP 5.5 deepTools plotFingerprint +// */ +// process MergedLibPlotFingerprint { +// tag "$name" +// label 'process_high' +// publishDir "${params.outdir}/bwa/mergedLibrary/deepTools/plotFingerprint", mode: 'copy' +// +// when: +// !params.skip_plot_fingerprint +// +// input: +// set val(name), file(bam) from ch_mlib_rm_orphan_bam_plotfingerprint +// +// output: +// file '*.{txt,pdf}' into ch_mlib_plotfingerprint_results +// file '*.raw.txt' into ch_mlib_plotfingerprint_mqc +// +// script: +// prefix = "${name}.mLb.clN" +// extend = (params.single_end && params.fragment_size > 0) ? "--extendReads ${params.fragment_size}" : '' +// """ +// plotFingerprint \\ +// --bamfiles ${bam[0]} \\ +// --plotFile ${prefix}.plotFingerprint.pdf \\ +// $extend \\ +// --labels $prefix \\ +// --outRawCounts ${prefix}.plotFingerprint.raw.txt \\ +// --outQualityMetrics ${prefix}.plotFingerprint.qcmetrics.txt \\ +// --skipZeros \\ +// --numberOfProcessors $task.cpus \\ +// --numberOfSamples $params.fingerprint_bins +// """ +// } +// +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// /* -- -- */ +// /* -- MERGE LIBRARY PEAK ANALYSIS -- */ +// /* -- -- */ +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// +// /* +// * STEP 6.1 Call peaks with MACS2 and calculate FRiP score +// */ +// process MergedLibMACSCallPeak { +// tag "$name" +// label 'process_medium' +// publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}", mode: 'copy', +// saveAs: { filename -> +// if (filename.endsWith(".tsv")) "qc/$filename" +// else if (filename.endsWith(".igv.txt")) null +// else filename +// } +// +// when: +// params.macs_gsize +// +// input: +// set val(name), file(bam), file(flagstat) from ch_mlib_rm_orphan_bam_macs.join(ch_mlib_rm_orphan_flagstat_macs, by: [0]) +// file mlib_peak_count_header from ch_mlib_peak_count_header +// file mlib_frip_score_header from ch_mlib_frip_score_header +// +// output: +// set val(name), file("*.{bed,xls,gappedPeak,bdg}") into ch_mlib_macs_output +// set val(name), file("*$PEAK_TYPE") into ch_mlib_macs_homer, +// ch_mlib_macs_qc, +// ch_mlib_macs_consensus, +// ch_mlib_macs_ataqv +// file "*igv.txt" into ch_mlib_macs_igv +// file "*_mqc.tsv" into ch_mlib_macs_mqc +// +// script: +// prefix = "${name}.mLb.clN" +// broad = params.narrow_peak ? '' : "--broad --broad-cutoff ${params.broad_cutoff}" +// format = params.single_end ? "BAM" : "BAMPE" +// pileup = params.save_macs_pileup ? "-B --SPMR" : "" +// """ +// macs2 callpeak \\ +// -t ${bam[0]} \\ +// $broad \\ +// -f $format \\ +// -g $params.macs_gsize \\ +// -n $prefix \\ +// $pileup \\ +// --keep-dup all \\ +// --nomodel +// +// cat ${prefix}_peaks.${PEAK_TYPE} | wc -l | awk -v OFS='\t' '{ print "${name}", \$1 }' | cat $mlib_peak_count_header - > ${prefix}_peaks.count_mqc.tsv +// +// READS_IN_PEAKS=\$(intersectBed -a ${bam[0]} -b ${prefix}_peaks.${PEAK_TYPE} -bed -c -f 0.20 | awk -F '\t' '{sum += \$NF} END {print sum}') +// grep 'mapped (' $flagstat | awk -v a="\$READS_IN_PEAKS" -v OFS='\t' '{print "${name}", a/\$1}' | cat $mlib_frip_score_header - > ${prefix}_peaks.FRiP_mqc.tsv +// +// find * -type f -name "*.${PEAK_TYPE}" -exec echo -e "bwa/mergedLibrary/macs/${PEAK_TYPE}/"{}"\\t0,0,178" \\; > ${prefix}_peaks.igv.txt +// """ +// } +// +// /* +// * STEP 6.2 Annotate peaks with HOMER +// */ +// process MergedLibAnnotatePeaks { +// tag "$name" +// label "process_medium" +// publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}", mode: 'copy' +// +// when: +// params.macs_gsize +// +// input: +// set val(name), file(peak) from ch_mlib_macs_homer +// file fasta from ch_fasta +// file gtf from ch_gtf +// +// output: +// file "*.txt" into ch_mlib_macs_annotate +// +// script: +// prefix = "${name}.mLb.clN" +// """ +// annotatePeaks.pl \\ +// $peak \\ +// $fasta \\ +// -gid \\ +// -gtf $gtf \\ +// -cpu $task.cpus \\ +// > ${prefix}_peaks.annotatePeaks.txt +// """ +// } +// +// /* +// * STEP 6.3 Aggregated QC plots for peaks, FRiP and peak-to-gene annotation +// */ +// process MergedLibPeakQC { +// label "process_medium" +// publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/qc", mode: 'copy' +// +// when: +// params.macs_gsize +// +// input: +// file peaks from ch_mlib_macs_qc.collect{ it[1] } +// file annos from ch_mlib_macs_annotate.collect() +// file mlib_peak_annotation_header from ch_mlib_peak_annotation_header +// +// output: +// file "*.{txt,pdf}" into ch_mlib_peak_qc +// file "*.tsv" into ch_mlib_peak_qc_mqc +// +// script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ +// suffix = 'mLb.clN' +// """ +// plot_macs_qc.r \\ +// -i ${peaks.join(',')} \\ +// -s ${peaks.join(',').replaceAll(".${suffix}_peaks.${PEAK_TYPE}","")} \\ +// -o ./ \\ +// -p macs_peak.${suffix} +// +// plot_homer_annotatepeaks.r \\ +// -i ${annos.join(',')} \\ +// -s ${annos.join(',').replaceAll(".${suffix}_peaks.annotatePeaks.txt","")} \\ +// -o ./ \\ +// -p macs_annotatePeaks.${suffix} +// +// cat $mlib_peak_annotation_header macs_annotatePeaks.${suffix}.summary.txt > macs_annotatePeaks.${suffix}.summary_mqc.tsv +// """ +// } +// +// /* +// * STEP 6.4 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection +// */ +// process MergedLibConsensusPeakSet { +// label 'process_long' +// publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: 'copy', +// saveAs: { filename -> +// if (filename.endsWith(".igv.txt")) null +// else filename +// } +// +// when: +// params.macs_gsize && (replicatesExist || multipleGroups) && !params.skip_consensus_peaks +// +// input: +// file peaks from ch_mlib_macs_consensus.collect{ it[1] } +// +// output: +// file "*.bed" into ch_mlib_macs_consensus_bed +// file "*.saf" into ch_mlib_macs_consensus_saf +// file "*.boolean.txt" into ch_mlib_macs_consensus_bool +// file "*.intersect.{txt,plot.pdf}" into ch_mlib_macs_consensus_intersect +// file "*igv.txt" into ch_mlib_macs_consensus_igv +// +// script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ +// suffix = 'mLb.clN' +// prefix = "consensus_peaks.${suffix}" +// mergecols = params.narrow_peak ? (2..10).join(',') : (2..9).join(',') +// collapsecols = params.narrow_peak ? (["collapse"]*9).join(',') : (["collapse"]*8).join(',') +// expandparam = params.narrow_peak ? "--is_narrow_peak" : "" +// """ +// sort -T '.' -k1,1 -k2,2n ${peaks.collect{it.toString()}.sort().join(' ')} \\ +// | mergeBed -c $mergecols -o $collapsecols > ${prefix}.txt +// +// macs2_merged_expand.py \\ +// ${prefix}.txt \\ +// ${peaks.collect{it.toString()}.sort().join(',').replaceAll("_peaks.${PEAK_TYPE}","")} \\ +// ${prefix}.boolean.txt \\ +// --min_replicates $params.min_reps_consensus \\ +// $expandparam +// +// awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$1, \$2, \$3, \$4, "0", "+" }' ${prefix}.boolean.txt > ${prefix}.bed +// +// echo -e "GeneID\tChr\tStart\tEnd\tStrand" > ${prefix}.saf +// awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$4, \$1, \$2, \$3, "+" }' ${prefix}.boolean.txt >> ${prefix}.saf +// +// sed -i 's/.${suffix}//g' ${prefix}.boolean.intersect.txt +// plot_peak_intersect.r -i ${prefix}.boolean.intersect.txt -o ${prefix}.boolean.intersect.plot.pdf +// +// find * -type f -name "${prefix}.bed" -exec echo -e "bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus/"{}"\\t0,0,0" \\; > ${prefix}.bed.igv.txt +// """ +// } +// +// /* +// * STEP 6.5 Annotate consensus peaks with HOMER, and add annotation to boolean output file +// */ +// process MergedLibConsensusPeakSetAnnotate { +// label "process_medium" +// publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: 'copy' +// +// when: +// params.macs_gsize && (replicatesExist || multipleGroups) && !params.skip_consensus_peaks +// +// input: +// file bed from ch_mlib_macs_consensus_bed +// file bool from ch_mlib_macs_consensus_bool +// file fasta from ch_fasta +// file gtf from ch_gtf +// +// output: +// file "*.annotatePeaks.txt" into ch_mlib_macs_consensus_annotate +// +// script: +// prefix = "consensus_peaks.mLb.clN" +// """ +// annotatePeaks.pl \\ +// $bed \\ +// $fasta \\ +// -gid \\ +// -gtf $gtf \\ +// -cpu $task.cpus \\ +// > ${prefix}.annotatePeaks.txt +// +// cut -f2- ${prefix}.annotatePeaks.txt | awk 'NR==1; NR > 1 {print \$0 | "sort -T '.' -k1,1 -k2,2n"}' | cut -f6- > tmp.txt +// paste $bool tmp.txt > ${prefix}.boolean.annotatePeaks.txt +// """ +// } +// +// /* +// * STEP 6.6 Count reads in consensus peaks with featureCounts and perform differential analysis with DESeq2 +// */ +// process MergedLibConsensusPeakSetDESeq { +// label 'process_medium' +// publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus/deseq2", mode: 'copy', +// saveAs: { filename -> +// if (filename.endsWith(".igv.txt")) null +// else filename +// } +// +// when: +// params.macs_gsize && replicatesExist && multipleGroups && !params.skip_consensus_peaks +// +// input: +// file bams from ch_mlib_name_bam_mlib_counts.collect{ it[1] } +// file saf from ch_mlib_macs_consensus_saf.collect() +// file mlib_deseq2_pca_header from ch_mlib_deseq2_pca_header +// file mlib_deseq2_clustering_header from ch_mlib_deseq2_clustering_header +// +// output: +// file "*featureCounts.txt" into ch_mlib_macs_consensus_counts +// file "*featureCounts.txt.summary" into ch_mlib_macs_consensus_counts_mqc +// file "*.{RData,results.txt,pdf,log}" into ch_mlib_macs_consensus_deseq_results +// file "sizeFactors" into ch_mlib_macs_consensus_deseq_factors +// file "*vs*/*.{pdf,txt}" into ch_mlib_macs_consensus_deseq_comp_results +// file "*vs*/*.bed" into ch_mlib_macs_consensus_deseq_comp_bed +// file "*igv.txt" into ch_mlib_macs_consensus_deseq_comp_igv +// file "*.tsv" into ch_mlib_macs_consensus_deseq_mqc +// +// script: +// prefix = "consensus_peaks.mLb.clN" +// bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() +// bam_ext = params.single_end ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" +// pe_params = params.single_end ? '' : "-p --donotsort" +// """ +// featureCounts \\ +// -F SAF \\ +// -O \\ +// --fracOverlap 0.2 \\ +// -T $task.cpus \\ +// $pe_params \\ +// -a $saf \\ +// -o ${prefix}.featureCounts.txt \\ +// ${bam_files.join(' ')} +// +// featurecounts_deseq2.r -i ${prefix}.featureCounts.txt -b '$bam_ext' -o ./ -p $prefix -s .mLb +// +// cat $mlib_deseq2_pca_header ${prefix}.pca.vals.txt > ${prefix}.pca.vals_mqc.tsv +// cat $mlib_deseq2_clustering_header ${prefix}.sample.dists.txt > ${prefix}.sample.dists_mqc.tsv +// +// find * -type f -name "*.FDR0.05.results.bed" -exec echo -e "bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus/deseq2/"{}"\\t255,0,0" \\; > ${prefix}.igv.txt +// """ +// } +// +// /* +// * STEP 6.7 Run ataqv on BAM file and corresponding peaks +// */ +// process MergedLibAtaqv { +// tag "$name" +// label 'process_medium' +// publishDir "${params.outdir}/bwa/mergedLibrary/ataqv/${PEAK_TYPE}", mode: 'copy' +// +// when: +// !params.skip_ataqv +// +// input: +// set val(name), file(bam), file(peak) from ch_mlib_bam_ataqv.join(ch_mlib_macs_ataqv, by: [0]) +// file autosomes from ch_genome_autosomes.collect() +// file tss_bed from ch_tss_bed +// +// output: +// file "*.json" into ch_mlib_ataqv +// +// script: +// peak_param = params.macs_gsize ? "--peak-file ${peak}" : "" +// mito_param = params.mito_name ? "--mitochondrial-reference-name ${params.mito_name}" : "" +// """ +// ataqv \\ +// --threads $task.cpus \\ +// $peak_param \\ +// --tss-file $tss_bed \\ +// --metrics-file ${name}.ataqv.json \\ +// --name $name \\ +// --ignore-read-groups \\ +// --autosomal-reference-file $autosomes \\ +// $mito_param \\ +// NA \\ +// ${bam[0]} +// """ +// } +// +// /* +// * STEP 6.8 run ataqv mkarv on all JSON files to render web app +// */ +// process MergedLibAtaqvMkarv { +// label 'process_medium' +// publishDir "${params.outdir}/bwa/mergedLibrary/ataqv/${PEAK_TYPE}", mode: 'copy' +// +// when: +// !params.skip_ataqv +// +// input: +// file json from ch_mlib_ataqv.collect() +// +// output: +// file "html" into ch_mlib_ataqv_mkarv +// +// script: +// """ +// mkarv \\ +// --concurrency $task.cpus \\ +// --force \\ +// ./html/ \\ +// ${json.join(' ')} +// """ +// } +// +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// /* -- -- */ +// /* -- MERGE REPLICATE BAM -- */ +// /* -- -- */ +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// +// /* +// * STEP 7 Merge library BAM files across all replicates +// */ +// ch_mlib_rm_orphan_bam_mrep +// .map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] } +// .groupTuple(by: [0]) +// .map { it -> [ it[0], it[1].flatten() ] } +// .set { ch_mlib_rm_orphan_bam_mrep } +// +// process MergedRepBAM { +// tag "$name" +// label 'process_medium' +// publishDir "${params.outdir}/bwa/mergedReplicate", mode: 'copy', +// saveAs: { filename -> +// if (filename.endsWith(".flagstat")) "samtools_stats/$filename" +// else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" +// else if (filename.endsWith(".stats")) "samtools_stats/$filename" +// else if (filename.endsWith(".metrics.txt")) "picard_metrics/$filename" +// else filename +// } +// +// input: +// set val(name), file(bams) from ch_mlib_rm_orphan_bam_mrep +// +// output: +// set val(name), file("*${prefix}.sorted.{bam,bam.bai}") into ch_mrep_bam_bigwig, +// ch_mrep_bam_macs +// set val(name), file("*.flagstat") into ch_mrep_bam_flagstat_bigwig, +// ch_mrep_bam_flagstat_macs, +// ch_mrep_bam_flagstat_mqc +// file "*.{idxstats,stats}" into ch_mrep_bam_stats_mqc +// file "*.txt" into ch_mrep_bam_metrics_mqc +// +// when: +// !params.skip_merge_replicates && replicatesExist +// +// script: +// prefix = "${name}.mRp.clN" +// bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() +// def avail_mem = 3 +// if (!task.memory) { +// log.info "[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." +// } else { +// avail_mem = task.memory.toGiga() +// } +// if (bam_files.size() > 1) { +// """ +// picard -Xmx${avail_mem}g MergeSamFiles \\ +// ${'INPUT='+bam_files.join(' INPUT=')} \\ +// OUTPUT=${name}.sorted.bam \\ +// SORT_ORDER=coordinate \\ +// VALIDATION_STRINGENCY=LENIENT \\ +// TMP_DIR=tmp +// samtools index ${name}.sorted.bam +// +// picard -Xmx${avail_mem}g MarkDuplicates \\ +// INPUT=${name}.sorted.bam \\ +// OUTPUT=${prefix}.sorted.bam \\ +// ASSUME_SORTED=true \\ +// REMOVE_DUPLICATES=true \\ +// METRICS_FILE=${prefix}.MarkDuplicates.metrics.txt \\ +// VALIDATION_STRINGENCY=LENIENT \\ +// TMP_DIR=tmp +// +// samtools index ${prefix}.sorted.bam +// samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat +// samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats +// samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats +// """ +// } else { +// """ +// ln -s ${bams[0]} ${prefix}.sorted.bam +// ln -s ${bams[1]} ${prefix}.sorted.bam.bai +// touch ${prefix}.MarkDuplicates.metrics.txt +// samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat +// samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats +// samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats +// """ +// } +// } +// +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// /* -- -- */ +// /* -- MERGE REPLICATE BAM POST-ANALYSIS -- */ +// /* -- -- */ +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// +// /* +// * STEP 8.1 Read depth normalised bigWig +// */ +// process MergedRepBigWig { +// tag "$name" +// label 'process_medium' +// publishDir "${params.outdir}/bwa/mergedReplicate/bigwig", mode: 'copy', +// saveAs: { filename -> +// if (filename.endsWith("scale_factor.txt")) "scale/$filename" +// else if (filename.endsWith(".bigWig")) "$filename" +// else null +// } +// +// when: +// !params.skip_merge_replicates && replicatesExist +// +// input: +// set val(name), file(bam), file(flagstat) from ch_mrep_bam_bigwig.join(ch_mrep_bam_flagstat_bigwig, by: [0]) +// file sizes from ch_genome_sizes_mrep_bigwig.collect() +// +// output: +// set val(name), file("*.bigWig") into ch_mrep_bigwig +// file "*scale_factor.txt" into ch_mrep_bigwig_scale +// file "*igv.txt" into ch_mrep_bigwig_igv +// +// script: +// prefix = "${name}.mRp.clN" +// pe_fragment = params.single_end ? "" : "-pc" +// extend = (params.single_end && params.fragment_size > 0) ? "-fs ${params.fragment_size}" : '' +// """ +// SCALE_FACTOR=\$(grep 'mapped (' $flagstat | awk '{print 1000000/\$1}') +// echo \$SCALE_FACTOR > ${prefix}.scale_factor.txt +// genomeCoverageBed -ibam ${bam[0]} -bg -scale \$SCALE_FACTOR $pe_fragment $extend | sort -T '.' -k1,1 -k2,2n > ${prefix}.bedGraph +// +// bedGraphToBigWig ${prefix}.bedGraph $sizes ${prefix}.bigWig +// +// find * -type f -name "*.bigWig" -exec echo -e "bwa/mergedReplicate/bigwig/"{}"\\t0,0,178" \\; > ${prefix}.bigWig.igv.txt +// """ +// } +// +// /* +// * STEP 8.2 Call peaks with MACS2 and calculate FRiP score +// */ +// process MergedRepMACSCallPeak { +// tag "$name" +// label 'process_medium' +// publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}", mode: 'copy', +// saveAs: { filename -> +// if (filename.endsWith(".tsv")) "qc/$filename" +// else if (filename.endsWith(".igv.txt")) null +// else filename +// } +// +// when: +// !params.skip_merge_replicates && replicatesExist && params.macs_gsize +// +// input: +// set val(name), file(bam), file(flagstat) from ch_mrep_bam_macs.join(ch_mrep_bam_flagstat_macs, by: [0]) +// file mrep_peak_count_header from ch_mrep_peak_count_header +// file mrep_frip_score_header from ch_mrep_frip_score_header +// +// output: +// set val(name), file("*.{bed,xls,gappedPeak,bdg}") into ch_mrep_macs_output +// set val(name), file("*$PEAK_TYPE") into ch_mrep_macs_homer, +// ch_mrep_macs_qc, +// ch_mrep_macs_consensus +// file "*igv.txt" into ch_mrep_macs_igv +// file "*_mqc.tsv" into ch_mrep_macs_mqc +// +// script: +// prefix = "${name}.mRp.clN" +// broad = params.narrow_peak ? '' : "--broad --broad-cutoff ${params.broad_cutoff}" +// format = params.single_end ? "BAM" : "BAMPE" +// pileup = params.save_macs_pileup ? "-B --SPMR" : "" +// """ +// macs2 callpeak \\ +// -t ${bam[0]} \\ +// $broad \\ +// -f $format \\ +// -g $params.macs_gsize \\ +// -n $prefix \\ +// $pileup \\ +// --keep-dup all \\ +// --nomodel +// +// cat ${prefix}_peaks.${PEAK_TYPE} | wc -l | awk -v OFS='\t' '{ print "${name}", \$1 }' | cat $mrep_peak_count_header - > ${prefix}_peaks.count_mqc.tsv +// +// READS_IN_PEAKS=\$(intersectBed -a ${bam[0]} -b ${prefix}_peaks.${PEAK_TYPE} -bed -c -f 0.20 | awk -F '\t' '{sum += \$NF} END {print sum}') +// grep 'mapped (' $flagstat | awk -v a="\$READS_IN_PEAKS" -v OFS='\t' '{print "${name}", a/\$1}' | cat $mrep_frip_score_header - > ${prefix}_peaks.FRiP_mqc.tsv +// +// find * -type f -name "*.${PEAK_TYPE}" -exec echo -e "bwa/mergedReplicate/macs/${PEAK_TYPE}/"{}"\\t0,0,178" \\; > ${prefix}_peaks.igv.txt +// """ +// } +// +// /* +// * STEP 8.3 Annotate peaks with HOMER +// */ +// process MergedRepAnnotatePeaks { +// tag "$name" +// label "process_medium" +// publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}", mode: 'copy' +// +// when: +// !params.skip_merge_replicates && replicatesExist && params.macs_gsize +// +// input: +// set val(name), file(peak) from ch_mrep_macs_homer +// file fasta from ch_fasta +// file gtf from ch_gtf +// +// output: +// file "*.txt" into ch_mrep_macs_annotate +// +// script: +// prefix = "${name}.mRp.clN" +// """ +// annotatePeaks.pl \\ +// $peak \\ +// $fasta \\ +// -gid \\ +// -gtf $gtf \\ +// -cpu $task.cpus \\ +// > ${prefix}_peaks.annotatePeaks.txt +// """ +// } +// +// /* +// * STEP 8.4 Aggregated QC plots for peaks, FRiP and peak-to-gene annotation +// */ +// process MergedRepPeakQC { +// label "process_medium" +// publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/qc", mode: 'copy' +// +// when: +// !params.skip_merge_replicates && replicatesExist && params.macs_gsize +// +// input: +// file peaks from ch_mrep_macs_qc.collect{ it[1] } +// file annos from ch_mrep_macs_annotate.collect() +// file mrep_peak_annotation_header from ch_mrep_peak_annotation_header +// +// output: +// file "*.{txt,pdf}" into ch_mrep_peak_qc +// file "*.tsv" into ch_mrep_peak_qc_mqc +// +// script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ +// suffix = 'mRp.clN' +// """ +// plot_macs_qc.r \\ +// -i ${peaks.join(',')} \\ +// -s ${peaks.join(',').replaceAll(".${suffix}_peaks.${PEAK_TYPE}","")} \\ +// -o ./ \\ +// -p macs_peak.${suffix} +// +// plot_homer_annotatepeaks.r \\ +// -i ${annos.join(',')} \\ +// -s ${annos.join(',').replaceAll(".${suffix}_peaks.annotatePeaks.txt","")} \\ +// -o ./ \\ +// -p macs_annotatePeaks.${suffix} +// +// cat $mrep_peak_annotation_header macs_annotatePeaks.${suffix}.summary.txt > macs_annotatePeaks.${suffix}.summary_mqc.tsv +// """ +// } +// +// /* +// * STEP 8.5 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection +// */ +// process MergedRepConsensusPeakSet { +// label 'process_long' +// publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: 'copy', +// saveAs: { filename -> +// if (filename.endsWith(".igv.txt")) null +// else filename +// } +// +// when: +// !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks +// +// input: +// file peaks from ch_mrep_macs_consensus.collect{ it[1] } +// +// output: +// file "*.bed" into ch_mrep_macs_consensus_bed +// file "*.saf" into ch_mrep_macs_consensus_saf +// file "*.boolean.txt" into ch_mrep_macs_consensus_bool +// file "*.intersect.{txt,plot.pdf}" into ch_mrep_macs_consensus_intersect +// file "*igv.txt" into ch_mrep_macs_consensus_igv +// +// script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ +// suffix = 'mRp.clN' +// prefix = "consensus_peaks.${suffix}" +// mergecols = params.narrow_peak ? (2..10).join(',') : (2..9).join(',') +// collapsecols = params.narrow_peak ? (["collapse"]*9).join(',') : (["collapse"]*8).join(',') +// expandparam = params.narrow_peak ? "--is_narrow_peak" : "" +// """ +// sort -T '.' -k1,1 -k2,2n ${peaks.collect{it.toString()}.sort().join(' ')} \\ +// | mergeBed -c $mergecols -o $collapsecols > ${prefix}.txt +// +// macs2_merged_expand.py \\ +// ${prefix}.txt \\ +// ${peaks.collect{it.toString()}.sort().join(',').replaceAll("_peaks.${PEAK_TYPE}","")} \\ +// ${prefix}.boolean.txt \\ +// $expandparam +// +// awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$1, \$2, \$3, \$4, "0", "+" }' ${prefix}.boolean.txt > ${prefix}.bed +// +// echo -e "GeneID\tChr\tStart\tEnd\tStrand" > ${prefix}.saf +// awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$4, \$1, \$2, \$3, "+" }' ${prefix}.boolean.txt >> ${prefix}.saf +// +// sed -i 's/.${suffix}//g' ${prefix}.boolean.intersect.txt +// plot_peak_intersect.r -i ${prefix}.boolean.intersect.txt -o ${prefix}.boolean.intersect.plot.pdf +// +// find * -type f -name "${prefix}.bed" -exec echo -e "bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus/"{}"\\t0,0,0" \\; > ${prefix}.bed.igv.txt +// """ +// } +// +// /* +// * STEP 8.6 Annotate consensus peaks with HOMER, and add annotation to boolean output file +// */ +// process MergedRepConsensusPeakSetAnnotate { +// label "process_medium" +// publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: 'copy' +// +// when: +// !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks +// +// input: +// file bed from ch_mrep_macs_consensus_bed +// file bool from ch_mrep_macs_consensus_bool +// file fasta from ch_fasta +// file gtf from ch_gtf +// +// output: +// file "*.annotatePeaks.txt" into ch_mrep_macs_consensus_annotate +// +// script: +// prefix = "consensus_peaks.mRp.clN" +// """ +// annotatePeaks.pl \\ +// $bed \\ +// $fasta \\ +// -gid \\ +// -gtf $gtf \\ +// -cpu $task.cpus \\ +// > ${prefix}.annotatePeaks.txt +// +// cut -f2- ${prefix}.annotatePeaks.txt | awk 'NR==1; NR > 1 {print \$0 | "sort -T '.' -k1,1 -k2,2n"}' | cut -f6- > tmp.txt +// paste $bool tmp.txt > ${prefix}.boolean.annotatePeaks.txt +// """ +// } +// +// /* +// * STEP 8.7 Count reads in consensus peaks with featureCounts and perform differential analysis with DESeq2 +// */ +// process MergedRepConsensusPeakSetDESeq { +// label 'process_medium' +// publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus/deseq2", mode: 'copy', +// saveAs: { filename -> +// if (filename.endsWith(".igv.txt")) null +// else filename +// } +// +// when: +// !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks +// +// input: +// file bams from ch_mlib_name_bam_mrep_counts.collect{ it[1] } +// file saf from ch_mrep_macs_consensus_saf.collect() +// file mrep_deseq2_pca_header from ch_mrep_deseq2_pca_header +// file mrep_deseq2_clustering_header from ch_mrep_deseq2_clustering_header +// +// output: +// file "*featureCounts.txt" into ch_mrep_macs_consensus_counts +// file "*featureCounts.txt.summary" into ch_mrep_macs_consensus_counts_mqc +// file "*.{RData,results.txt,pdf,log}" into ch_mrep_macs_consensus_deseq_results +// file "sizeFactors" into ch_mrep_macs_consensus_deseq_factors +// file "*vs*/*.{pdf,txt}" into ch_mrep_macs_consensus_deseq_comp_results +// file "*vs*/*.bed" into ch_mrep_macs_consensus_deseq_comp_bed +// file "*igv.txt" into ch_mrep_macs_consensus_deseq_comp_igv +// file "*.tsv" into ch_mrep_macs_consensus_deseq_mqc +// +// script: +// prefix = "consensus_peaks.mRp.clN" +// bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() +// bam_ext = params.single_end ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" +// pe_params = params.single_end ? '' : "-p --donotsort" +// """ +// featureCounts \\ +// -F SAF \\ +// -O \\ +// --fracOverlap 0.2 \\ +// -T $task.cpus \\ +// $pe_params \\ +// -a $saf \\ +// -o ${prefix}.featureCounts.txt \\ +// ${bam_files.join(' ')} +// +// featurecounts_deseq2.r -i ${prefix}.featureCounts.txt -b '$bam_ext' -o ./ -p $prefix -s .mLb +// +// cat $mrep_deseq2_pca_header ${prefix}.pca.vals.txt > ${prefix}.pca.vals_mqc.tsv +// cat $mrep_deseq2_clustering_header ${prefix}.sample.dists.txt > ${prefix}.sample.dists_mqc.tsv +// +// find * -type f -name "*.FDR0.05.results.bed" -exec echo -e "bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus/deseq2/"{}"\\t255,0,0" \\; > ${prefix}.igv.txt +// """ +// } +// +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// /* -- -- */ +// /* -- IGV -- */ +// /* -- -- */ +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// +// /* +// * STEP 9 - Create IGV session file +// */ +// process IGV { +// publishDir "${params.outdir}/igv/${PEAK_TYPE}", mode: 'copy' +// +// when: +// !params.skip_igv +// +// input: +// file fasta from ch_fasta +// +// file bigwigs from ch_mlib_bigwig_igv.collect().ifEmpty([]) +// file peaks from ch_mlib_macs_igv.collect().ifEmpty([]) +// file consensus_peaks from ch_mlib_macs_consensus_igv.collect().ifEmpty([]) +// file differential_peaks from ch_mlib_macs_consensus_deseq_comp_igv.collect().ifEmpty([]) +// +// file rbigwigs from ch_mrep_bigwig_igv.collect().ifEmpty([]) +// file rpeaks from ch_mrep_macs_igv.collect().ifEmpty([]) +// file rconsensus_peaks from ch_mrep_macs_consensus_igv.collect().ifEmpty([]) +// file rdifferential_peaks from ch_mrep_macs_consensus_deseq_comp_igv.collect().ifEmpty([]) +// +// output: +// file "*.{txt,xml}" into ch_igv_session +// +// script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ +// """ +// cat *.txt > igv_files.txt +// igv_files_to_session.py igv_session.xml igv_files.txt ../../reference_genome/${fasta.getName()} --path_prefix '../../' +// """ +// } +// +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// /* -- -- */ +// /* -- MULTIQC -- */ +// /* -- -- */ +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// +// /* +// * Parse software version numbers +// */ +// process get_software_versions { +// publishDir "${params.outdir}/pipeline_info", mode: 'copy', +// saveAs: { filename -> +// if (filename.indexOf(".csv") > 0) filename +// else null +// } +// +// output: +// file 'software_versions_mqc.yaml' into ch_software_versions_mqc +// file "software_versions.csv" +// +// script: +// """ +// echo $workflow.manifest.version > v_pipeline.txt +// echo $workflow.nextflow.version > v_nextflow.txt +// fastqc --version > v_fastqc.txt +// trim_galore --version > v_trim_galore.txt +// echo \$(bwa 2>&1) > v_bwa.txt +// samtools --version > v_samtools.txt +// bedtools --version > v_bedtools.txt +// echo \$(bamtools --version 2>&1) > v_bamtools.txt +// echo \$(plotFingerprint --version 2>&1) > v_deeptools.txt || true +// picard MarkDuplicates --version &> v_picard.txt || true +// echo \$(R --version 2>&1) > v_R.txt +// python -c "import pysam; print(pysam.__version__)" > v_pysam.txt +// echo \$(macs2 --version 2>&1) > v_macs2.txt +// touch v_homer.txt +// echo \$(ataqv --version 2>&1) > v_ataqv.txt +// echo \$(featureCounts -v 2>&1) > v_featurecounts.txt +// preseq &> v_preseq.txt +// multiqc --version > v_multiqc.txt +// scrape_software_versions.py &> software_versions_mqc.yaml +// """ +// } +// +// def create_workflow_summary(summary) { +// +// def yaml_file = workDir.resolve('workflow_summary_mqc.yaml') +// yaml_file.text = """ +// id: 'nf-core-atacseq-summary' +// description: " - this information is collected when the pipeline is started." +// section_name: 'nf-core/atacseq Workflow Summary' +// section_href: 'https://github.com/nf-core/atacseq' +// plot_type: 'html' +// data: | +//
+// ${summary.collect { k,v -> "
$k
${v ?: 'N/A'}
" }.join("\n")} +//
+// """.stripIndent() +// +// return yaml_file +// } +// +// /* +// * STEP 10 - MultiQC +// */ +// process MultiQC { +// publishDir "${params.outdir}/multiqc/${PEAK_TYPE}", mode: 'copy' +// +// when: +// !params.skip_multiqc +// +// input: +// file multiqc_config from ch_multiqc_config +// +// file ('software_versions/*') from ch_software_versions_mqc.collect() +// file ('workflow_summary/*') from create_workflow_summary(summary) +// +// file ('fastqc/*') from ch_fastqc_reports_mqc.collect().ifEmpty([]) +// file ('trimgalore/*') from ch_trimgalore_results_mqc.collect().ifEmpty([]) +// file ('trimgalore/fastqc/*') from ch_trimgalore_fastqc_reports_mqc.collect().ifEmpty([]) +// +// file ('alignment/library/*') from ch_sort_bam_flagstat_mqc.collect() +// +// file ('alignment/mergedLibrary/*') from ch_mlib_bam_stats_mqc.collect() +// file ('alignment/mergedLibrary/*') from ch_mlib_rm_orphan_flagstat_mqc.collect{it[1]} +// file ('alignment/mergedLibrary/*') from ch_mlib_rm_orphan_stats_mqc.collect() +// file ('alignment/mergedLibrary/picard_metrics/*') from ch_mlib_bam_metrics_mqc.collect() +// file ('alignment/mergedLibrary/picard_metrics/*') from ch_mlib_collectmetrics_mqc.collect() +// file ('macs/mergedLibrary/*') from ch_mlib_macs_mqc.collect().ifEmpty([]) +// file ('macs/mergedLibrary/*') from ch_mlib_peak_qc_mqc.collect().ifEmpty([]) +// file ('macs/mergedLibrary/consensus/*') from ch_mlib_macs_consensus_counts_mqc.collect().ifEmpty([]) +// file ('macs/mergedLibrary/consensus/*') from ch_mlib_macs_consensus_deseq_mqc.collect().ifEmpty([]) +// file ('preseq/*') from ch_mlib_preseq_mqc.collect().ifEmpty([]) +// file ('deeptools/*') from ch_mlib_plotprofile_mqc.collect().ifEmpty([]) +// file ('deeptools/*') from ch_mlib_plotfingerprint_mqc.collect().ifEmpty([]) +// +// file ('alignment/mergedReplicate/*') from ch_mrep_bam_flagstat_mqc.collect{it[1]}.ifEmpty([]) +// file ('alignment/mergedReplicate/*') from ch_mrep_bam_stats_mqc.collect().ifEmpty([]) +// file ('alignment/mergedReplicate/*') from ch_mrep_bam_metrics_mqc.collect().ifEmpty([]) +// file ('macs/mergedReplicate/*') from ch_mrep_macs_mqc.collect().ifEmpty([]) +// file ('macs/mergedReplicate/*') from ch_mrep_peak_qc_mqc.collect().ifEmpty([]) +// file ('macs/mergedReplicate/consensus/*') from ch_mrep_macs_consensus_counts_mqc.collect().ifEmpty([]) +// file ('macs/mergedReplicate/consensus/*') from ch_mrep_macs_consensus_deseq_mqc.collect().ifEmpty([]) +// +// output: +// file "*multiqc_report.html" into ch_multiqc_report +// file "*_data" +// file "multiqc_plots" +// +// script: +// rtitle = custom_runName ? "--title \"$custom_runName\"" : '' +// rfilename = custom_runName ? "--filename " + custom_runName.replaceAll('\\W','_').replaceAll('_+','_') + "_multiqc_report" : '' +// """ +// multiqc . -f $rtitle $rfilename --config $multiqc_config \\ +// -m custom_content -m fastqc -m cutadapt -m samtools -m picard -m preseq -m featureCounts -m deeptools +// """ +// } +// +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// /* -- -- */ +// /* -- REPORTS/DOCUMENTATION -- */ +// /* -- -- */ +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// +// /* +// * STEP 11 - Output description HTML +// */ +// process output_documentation { +// publishDir "${params.outdir}/Documentation", mode: 'copy' +// +// input: +// file output_docs from ch_output_docs +// +// output: +// file "results_description.html" +// +// script: +// """ +// markdown_to_html.r $output_docs results_description.html +// """ +// } +// +// /* +// * Completion e-mail notification +// */ +// workflow.onComplete { +// +// // Set up the e-mail variables +// def subject = "[nf-core/atacseq] Successful: $workflow.runName" +// if (!workflow.success) { +// subject = "[nf-core/atacseq] FAILED: $workflow.runName" +// } +// def email_fields = [:] +// email_fields['version'] = workflow.manifest.version +// email_fields['runName'] = custom_runName ?: workflow.runName +// email_fields['success'] = workflow.success +// email_fields['dateComplete'] = workflow.complete +// email_fields['duration'] = workflow.duration +// email_fields['exitStatus'] = workflow.exitStatus +// email_fields['errorMessage'] = (workflow.errorMessage ?: 'None') +// email_fields['errorReport'] = (workflow.errorReport ?: 'None') +// email_fields['commandLine'] = workflow.commandLine +// email_fields['projectDir'] = workflow.projectDir +// email_fields['summary'] = summary +// email_fields['summary']['Date Started'] = workflow.start +// email_fields['summary']['Date Completed'] = workflow.complete +// email_fields['summary']['Pipeline script file path'] = workflow.scriptFile +// email_fields['summary']['Pipeline script hash ID'] = workflow.scriptId +// if (workflow.repository) email_fields['summary']['Pipeline repository Git URL'] = workflow.repository +// if (workflow.commitId) email_fields['summary']['Pipeline repository Git Commit'] = workflow.commitId +// if (workflow.revision) email_fields['summary']['Pipeline Git branch/tag'] = workflow.revision +// if (workflow.container) email_fields['summary']['Docker image'] = workflow.container +// email_fields['summary']['Nextflow Version'] = workflow.nextflow.version +// email_fields['summary']['Nextflow Build'] = workflow.nextflow.build +// email_fields['summary']['Nextflow Compile Timestamp'] = workflow.nextflow.timestamp +// +// // On success try attach the multiqc report +// def mqc_report = null +// try { +// if (workflow.success) { +// mqc_report = ch_multiqc_report.getVal() +// if (mqc_report.getClass() == ArrayList) { +// log.warn "[nf-core/atacseq] Found multiple reports from process 'multiqc', will use only one" +// mqc_report = mqc_report[0] +// } +// } +// } catch (all) { +// log.warn "[nf-core/atacseq] Could not attach MultiQC report to summary email" +// } +// +// // Check if we are only sending emails on failure +// email_address = params.email +// if (!params.email && params.email_on_fail && !workflow.success) { +// email_address = params.email_on_fail +// } +// +// // Render the TXT template +// def engine = new groovy.text.GStringTemplateEngine() +// def tf = new File("$baseDir/assets/email_template.txt") +// def txt_template = engine.createTemplate(tf).make(email_fields) +// def email_txt = txt_template.toString() +// +// // Render the HTML template +// def hf = new File("$baseDir/assets/email_template.html") +// def html_template = engine.createTemplate(hf).make(email_fields) +// def email_html = html_template.toString() +// +// // Render the sendmail template +// def smail_fields = [ email: email_address, subject: subject, email_txt: email_txt, email_html: email_html, baseDir: "$baseDir", mqcFile: mqc_report, mqcMaxSize: params.max_multiqc_email_size.toBytes() ] +// def sf = new File("$baseDir/assets/sendmail_template.txt") +// def sendmail_template = engine.createTemplate(sf).make(smail_fields) +// def sendmail_html = sendmail_template.toString() +// +// // Send the HTML e-mail +// if (email_address) { +// try { +// if (params.plaintext_email) { throw GroovyException('Send plaintext e-mail, not HTML') } +// // Try to send HTML e-mail using sendmail +// [ 'sendmail', '-t' ].execute() << sendmail_html +// log.info "[nf-core/atacseq] Sent summary e-mail to $email_address (sendmail)" +// } catch (all) { +// // Catch failures and try with plaintext +// [ 'mail', '-s', subject, email_address ].execute() << email_txt +// log.info "[nf-core/atacseq] Sent summary e-mail to $email_address (mail)" +// } +// } +// +// // Write summary e-mail HTML to a file +// def output_d = new File("${params.outdir}/pipeline_info/") +// if (!output_d.exists()) { +// output_d.mkdirs() +// } +// def output_hf = new File(output_d, "pipeline_report.html") +// output_hf.withWriter { w -> w << email_html } +// def output_tf = new File(output_d, "pipeline_report.txt") +// output_tf.withWriter { w -> w << email_txt } +// +// c_reset = params.monochrome_logs ? '' : "\033[0m"; +// c_purple = params.monochrome_logs ? '' : "\033[0;35m"; +// c_green = params.monochrome_logs ? '' : "\033[0;32m"; +// c_red = params.monochrome_logs ? '' : "\033[0;31m"; +// +// if (workflow.stats.ignoredCount > 0 && workflow.success) { +// log.info "${c_purple}Warning, pipeline completed, but with errored process(es) ${c_reset}" +// log.info "${c_red}Number of ignored errored process(es) : ${workflow.stats.ignoredCount} ${c_reset}" +// log.info "${c_green}Number of successfully ran process(es) : ${workflow.stats.succeedCount} ${c_reset}" +// } +// +// if (workflow.success) { +// log.info "${c_purple}[nf-core/atacseq]${c_green} Pipeline completed successfully${c_reset}" +// } else { +// checkHostname() +// log.info "${c_purple}[nf-core/atacseq]${c_red} Pipeline completed with errors${c_reset}" +// } +// +// } /////////////////////////////////////////////////////////////////////////////// /////////////////////////////////////////////////////////////////////////////// From 1054c920b5cd0a7b32675184bb67c4ef79d9833a Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 30 Jan 2020 13:54:30 +0000 Subject: [PATCH 079/226] Add README.txt description --- docs/output.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/docs/output.md b/docs/output.md index c4c2f6fc..6ad1b844 100755 --- a/docs/output.md +++ b/docs/output.md @@ -318,7 +318,7 @@ You can skip this portion of the analysis by specifying the `--skip_merge_replic *Output directories*: * `reference_genome/` - A number of genome-specific files are generated by the pipeline in order to aid in the filtering of the data, and because they are required by standard tools such as BEDTools. These can be found in this directory along with the genome fasta file which is required by IGV. + A number of genome-specific files are generated by the pipeline in order to aid in the filtering of the data, and because they are required by standard tools such as BEDTools. These can be found in this directory along with the genome fasta file which is required by IGV. If using a genome from AWS iGenomes and if it exists a `README.txt` file containing information about the annotation version will also be saved in this directory. * `reference_genome/BWAIndex/` If the `--save_reference` parameter is provided then the alignment indices generated by the pipeline will be saved in this directory. This can be quite a time-consuming process so it permits their reuse for future runs of the pipeline or for other purposes. From bff3d8b31fb6ec48f4e849753c99b1ae57403db8 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 30 Jan 2020 14:01:51 +0000 Subject: [PATCH 080/226] Update CHANGELOG --- CHANGELOG.md | 5 +++-- 1 file changed, 3 insertions(+), 2 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index c8d88d88..f727f28c 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -10,12 +10,13 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Added` * [#63](https://github.com/nf-core/atacseq/issues/63) - Added multicore support for Trim Galore! +* [#75](https://github.com/nf-core/atacseq/issues/75) - Include gene annotation versions in multiqc report * Added GitHub Actions CI tests ### `Fixed` -* [#118](https://github.com/nf-core/chipseq/issues/118) - Running on with SGE -* [#132](https://github.com/nf-core/chipseq/issues/132) - BigWig Error: sort: cannot create temporary file in '': Read-only file system +* [nf-core/chipseq#118](https://github.com/nf-core/chipseq/issues/118) - Running on with SGE +* [nf-core/chipseq#132](https://github.com/nf-core/chipseq/issues/132) - BigWig Error: sort: cannot create temporary file in '': Read-only file system * [#73](https://github.com/nf-core/atacseq/issues/73) - macs_annotatePeaks.mLb.clN.summary.txt file is not created * Make executables in `bin/` compatible with Python 3 From 8f9c883ae33e9257839172d10761f145b28292f2 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 30 Jan 2020 17:52:09 +0000 Subject: [PATCH 081/226] Comment in code --- main.nf | 3252 +++++++++++++++++++++++++++---------------------------- 1 file changed, 1626 insertions(+), 1626 deletions(-) diff --git a/main.nf b/main.nf index fc7dd155..e6294f73 100644 --- a/main.nf +++ b/main.nf @@ -469,1632 +469,1632 @@ process MakeGenomeFilter { """ } -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// /* -- -- */ -// /* -- FASTQ QC -- */ -// /* -- -- */ -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// -// /* -// * STEP 1 - FastQC -// */ -// process FastQC { -// tag "$name" -// label 'process_medium' -// publishDir "${params.outdir}/fastqc", mode: 'copy', -// saveAs: { filename -> -// filename.endsWith(".zip") ? "zips/$filename" : "$filename" -// } -// -// when: -// !params.skip_fastqc -// -// input: -// set val(name), file(reads) from ch_raw_reads_fastqc -// -// output: -// file "*.{zip,html}" into ch_fastqc_reports_mqc -// -// script: -// // Added soft-links to original fastqs for consistent naming in MultiQC -// if (params.single_end) { -// """ -// [ ! -f ${name}.fastq.gz ] && ln -s $reads ${name}.fastq.gz -// fastqc -q -t $task.cpus ${name}.fastq.gz -// """ -// } else { -// """ -// [ ! -f ${name}_1.fastq.gz ] && ln -s ${reads[0]} ${name}_1.fastq.gz -// [ ! -f ${name}_2.fastq.gz ] && ln -s ${reads[1]} ${name}_2.fastq.gz -// fastqc -q -t $task.cpus ${name}_1.fastq.gz -// fastqc -q -t $task.cpus ${name}_2.fastq.gz -// """ -// } -// } -// -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// /* -- -- */ -// /* -- ADAPTER TRIMMING -- */ -// /* -- -- */ -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// -// /* -// * STEP 2 - Trim Galore! -// */ -// if (params.skip_trimming) { -// ch_trimmed_reads = ch_raw_reads_trimgalore -// ch_trimgalore_results_mqc = Channel.empty() -// ch_trimgalore_fastqc_reports_mqc = Channel.empty() -// } else { -// process TrimGalore { -// tag "$name" -// label 'process_high' -// publishDir "${params.outdir}/trim_galore", mode: 'copy', -// saveAs: { filename -> -// if (filename.endsWith(".html")) "fastqc/$filename" -// else if (filename.endsWith(".zip")) "fastqc/zips/$filename" -// else if (filename.endsWith("trimming_report.txt")) "logs/$filename" -// else params.save_trimmed ? filename : null -// } -// -// input: -// set val(name), file(reads) from ch_raw_reads_trimgalore -// -// output: -// set val(name), file("*.fq.gz") into ch_trimmed_reads -// file "*.txt" into ch_trimgalore_results_mqc -// file "*.{zip,html}" into ch_trimgalore_fastqc_reports_mqc -// -// script: -// // Calculate number of --cores for TrimGalore based on value of task.cpus -// // See: https://github.com/FelixKrueger/TrimGalore/blob/master/Changelog.md#version-060-release-on-1-mar-2019 -// // See: https://github.com/nf-core/atacseq/pull/65 -// def cores = 1 -// if (task.cpus) { -// cores = (task.cpus as int) - 4 -// if (params.single_end) cores = (task.cpus as int) - 3 -// if (cores < 1) cores = 1 -// if (cores > 4) cores = 4 -// } -// -// // Added soft-links to original fastqs for consistent naming in MultiQC -// c_r1 = params.clip_r1 > 0 ? "--clip_r1 ${params.clip_r1}" : '' -// c_r2 = params.clip_r2 > 0 ? "--clip_r2 ${params.clip_r2}" : '' -// tpc_r1 = params.three_prime_clip_r1 > 0 ? "--three_prime_clip_r1 ${params.three_prime_clip_r1}" : '' -// tpc_r2 = params.three_prime_clip_r2 > 0 ? "--three_prime_clip_r2 ${params.three_prime_clip_r2}" : '' -// nextseq = params.trim_nextseq > 0 ? "--nextseq ${params.trim_nextseq}" : '' -// -// // Added soft-links to original fastqs for consistent naming in MultiQC -// if (params.single_end) { -// """ -// [ ! -f ${name}.fastq.gz ] && ln -s $reads ${name}.fastq.gz -// trim_galore --cores $cores --fastqc --gzip $c_r1 $tpc_r1 $nextseq ${name}.fastq.gz -// """ -// } else { -// """ -// [ ! -f ${name}_1.fastq.gz ] && ln -s ${reads[0]} ${name}_1.fastq.gz -// [ ! -f ${name}_2.fastq.gz ] && ln -s ${reads[1]} ${name}_2.fastq.gz -// trim_galore --cores $cores --paired --fastqc --gzip $c_r1 $c_r2 $tpc_r1 $tpc_r2 $nextseq ${name}_1.fastq.gz ${name}_2.fastq.gz -// """ -// } -// } -// } -// -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// /* -- -- */ -// /* -- ALIGN -- */ -// /* -- -- */ -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// -// /* -// * STEP 3.1 - Align read 1 with bwa -// */ -// process BWAMem { -// tag "$name" -// label 'process_high' -// -// input: -// set val(name), file(reads) from ch_trimmed_reads -// file index from ch_bwa_index.collect() -// -// output: -// set val(name), file("*.bam") into ch_bwa_bam -// -// script: -// prefix = "${name}.Lb" -// rg = "\'@RG\\tID:${name}\\tSM:${name.split('_')[0..-2].join('_')}\\tPL:ILLUMINA\\tLB:${name}\\tPU:1\'" -// if (params.seq_center) { -// rg = "\'@RG\\tID:${name}\\tSM:${name.split('_')[0..-2].join('_')}\\tPL:ILLUMINA\\tLB:${name}\\tPU:1\\tCN:${params.seq_center}\'" -// } -// """ -// bwa mem \\ -// -t $task.cpus \\ -// -M \\ -// -R $rg \\ -// ${index}/${bwa_base} \\ -// $reads \\ -// | samtools view -@ $task.cpus -b -h -F 0x0100 -O BAM -o ${prefix}.bam - -// """ -// } -// -// /* -// * STEP 3.2 - Convert .bam to coordinate sorted .bam -// */ -// process SortBAM { -// tag "$name" -// label 'process_medium' -// if (params.save_align_intermeds) { -// publishDir path: "${params.outdir}/bwa/library", mode: 'copy', -// saveAs: { filename -> -// if (filename.endsWith(".flagstat")) "samtools_stats/$filename" -// else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" -// else if (filename.endsWith(".stats")) "samtools_stats/$filename" -// else filename -// } -// } -// -// input: -// set val(name), file(bam) from ch_bwa_bam -// -// output: -// set val(name), file("*.sorted.{bam,bam.bai}") into ch_sort_bam_merge -// file "*.{flagstat,idxstats,stats}" into ch_sort_bam_flagstat_mqc -// -// script: -// prefix = "${name}.Lb" -// """ -// samtools sort -@ $task.cpus -o ${prefix}.sorted.bam -T $name $bam -// samtools index ${prefix}.sorted.bam -// samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat -// samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats -// samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats -// """ -// } -// -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// /* -- -- */ -// /* -- MERGE LIBRARY BAM -- */ -// /* -- -- */ -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// -// /* -// * STEP 4.1 Merge BAM files for all libraries from same replicate -// */ -// ch_sort_bam_merge -// .map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] } -// .groupTuple(by: [0]) -// .map { it -> [ it[0], it[1].flatten() ] } -// .set { ch_sort_bam_merge } -// -// process MergedLibBAM { -// tag "$name" -// label 'process_medium' -// publishDir "${params.outdir}/bwa/mergedLibrary", mode: 'copy', -// saveAs: { filename -> -// if (filename.endsWith(".flagstat")) "samtools_stats/$filename" -// else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" -// else if (filename.endsWith(".stats")) "samtools_stats/$filename" -// else if (filename.endsWith(".metrics.txt")) "picard_metrics/$filename" -// else params.save_align_intermeds ? filename : null -// } -// -// input: -// set val(name), file(bams) from ch_sort_bam_merge -// -// output: -// set val(name), file("*${prefix}.sorted.{bam,bam.bai}") into ch_mlib_bam_filter, -// ch_mlib_bam_preseq, -// ch_mlib_bam_ataqv -// file "*.{flagstat,idxstats,stats}" into ch_mlib_bam_stats_mqc -// file "*.txt" into ch_mlib_bam_metrics_mqc -// -// script: -// prefix = "${name}.mLb.mkD" -// bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() -// def avail_mem = 3 -// if (!task.memory) { -// log.info "[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." -// } else { -// avail_mem = task.memory.toGiga() -// } -// if (bam_files.size() > 1) { -// """ -// picard -Xmx${avail_mem}g MergeSamFiles \\ -// ${'INPUT='+bam_files.join(' INPUT=')} \\ -// OUTPUT=${name}.sorted.bam \\ -// SORT_ORDER=coordinate \\ -// VALIDATION_STRINGENCY=LENIENT \\ -// TMP_DIR=tmp -// samtools index ${name}.sorted.bam -// -// picard -Xmx${avail_mem}g MarkDuplicates \\ -// INPUT=${name}.sorted.bam \\ -// OUTPUT=${prefix}.sorted.bam \\ -// ASSUME_SORTED=true \\ -// REMOVE_DUPLICATES=false \\ -// METRICS_FILE=${prefix}.MarkDuplicates.metrics.txt \\ -// VALIDATION_STRINGENCY=LENIENT \\ -// TMP_DIR=tmp -// -// samtools index ${prefix}.sorted.bam -// samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats -// samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat -// samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats -// """ -// } else { -// """ -// picard -Xmx${avail_mem}g MarkDuplicates \\ -// INPUT=${bam_files[0]} \\ -// OUTPUT=${prefix}.sorted.bam \\ -// ASSUME_SORTED=true \\ -// REMOVE_DUPLICATES=false \\ -// METRICS_FILE=${prefix}.MarkDuplicates.metrics.txt \\ -// VALIDATION_STRINGENCY=LENIENT \\ -// TMP_DIR=tmp -// -// samtools index ${prefix}.sorted.bam -// samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats -// samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat -// samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats -// """ -// } -// } -// -// /* -// * STEP 4.2 Filter BAM file at merged library-level -// */ -// process MergedLibBAMFilter { -// tag "$name" -// label 'process_medium' -// publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', -// saveAs: { filename -> -// if (params.single_end || params.save_align_intermeds) { -// if (filename.endsWith(".flagstat")) "samtools_stats/$filename" -// else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" -// else if (filename.endsWith(".stats")) "samtools_stats/$filename" -// else if (filename.endsWith(".sorted.bam")) filename -// else if (filename.endsWith(".sorted.bam.bai")) filename -// else null -// } -// } -// -// input: -// set val(name), file(bam) from ch_mlib_bam_filter -// file bed from ch_genome_filter_regions.collect() -// file bamtools_filter_config from ch_bamtools_filter_config -// -// output: -// set val(name), file("*.{bam,bam.bai}") into ch_mlib_filter_bam -// set val(name), file("*.flagstat") into ch_mlib_filter_bam_flagstat -// file "*.{idxstats,stats}" into ch_mlib_filter_bam_stats_mqc -// -// script: -// prefix = params.single_end ? "${name}.mLb.clN" : "${name}.mLb.flT" -// filter_params = params.single_end ? "-F 0x004" : "-F 0x004 -F 0x0008 -f 0x001" -// dup_params = params.keep_dups ? "" : "-F 0x0400" -// multimap_params = params.keep_multi_map ? "" : "-q 1" -// blacklist_params = params.blacklist ? "-L $bed" : "" -// name_sort_bam = params.single_end ? "" : "samtools sort -n -@ $task.cpus -o ${prefix}.bam -T $prefix ${prefix}.sorted.bam" -// """ -// samtools view \\ -// $filter_params \\ -// $dup_params \\ -// $multimap_params \\ -// $blacklist_params \\ -// -b ${bam[0]} \\ -// | bamtools filter \\ -// -out ${prefix}.sorted.bam \\ -// -script $bamtools_filter_config -// -// samtools index ${prefix}.sorted.bam -// samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat -// samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats -// samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats -// -// $name_sort_bam -// """ -// } -// -// /* -// * STEP 4.3 Remove orphan reads from paired-end BAM file -// */ -// if (params.single_end) { -// ch_mlib_filter_bam -// .into { ch_mlib_rm_orphan_bam_metrics; -// ch_mlib_rm_orphan_bam_bigwig; -// ch_mlib_rm_orphan_bam_macs; -// ch_mlib_rm_orphan_bam_plotfingerprint; -// ch_mlib_rm_orphan_bam_mrep; -// ch_mlib_name_bam_mlib_counts; -// ch_mlib_name_bam_mrep_counts } -// -// ch_mlib_filter_bam_flagstat -// .into { ch_mlib_rm_orphan_flagstat_bigwig; -// ch_mlib_rm_orphan_flagstat_macs; -// ch_mlib_rm_orphan_flagstat_mqc } -// -// ch_mlib_filter_bam_stats_mqc -// .set { ch_mlib_rm_orphan_stats_mqc } -// } else { -// process MergedLibBAMRemoveOrphan { -// tag "$name" -// label 'process_medium' -// publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', -// saveAs: { filename -> -// if (filename.endsWith(".flagstat")) "samtools_stats/$filename" -// else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" -// else if (filename.endsWith(".stats")) "samtools_stats/$filename" -// else if (filename.endsWith(".sorted.bam")) filename -// else if (filename.endsWith(".sorted.bam.bai")) filename -// else null -// } -// -// input: -// set val(name), file(bam) from ch_mlib_filter_bam -// -// output: -// set val(name), file("*.sorted.{bam,bam.bai}") into ch_mlib_rm_orphan_bam_metrics, -// ch_mlib_rm_orphan_bam_bigwig, -// ch_mlib_rm_orphan_bam_macs, -// ch_mlib_rm_orphan_bam_plotfingerprint, -// ch_mlib_rm_orphan_bam_mrep -// set val(name), file("${prefix}.bam") into ch_mlib_name_bam_mlib_counts, -// ch_mlib_name_bam_mrep_counts -// set val(name), file("*.flagstat") into ch_mlib_rm_orphan_flagstat_bigwig, -// ch_mlib_rm_orphan_flagstat_macs, -// ch_mlib_rm_orphan_flagstat_mqc -// file "*.{idxstats,stats}" into ch_mlib_rm_orphan_stats_mqc -// -// script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ -// prefix = "${name}.mLb.clN" -// """ -// bampe_rm_orphan.py ${bam[0]} ${prefix}.bam --only_fr_pairs -// -// samtools sort -@ $task.cpus -o ${prefix}.sorted.bam -T $prefix ${prefix}.bam -// samtools index ${prefix}.sorted.bam -// samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat -// samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats -// samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats -// """ -// } -// } -// -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// /* -- -- */ -// /* -- MERGE LIBRARY BAM POST-ANALYSIS -- */ -// /* -- -- */ -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// -// /* -// * STEP 5.1 preseq analysis after merging libraries and before filtering -// */ -// process MergedLibPreseq { -// tag "$name" -// label 'process_low' -// publishDir "${params.outdir}/bwa/mergedLibrary/preseq", mode: 'copy' -// -// when: -// !params.skip_preseq -// -// input: -// set val(name), file(bam) from ch_mlib_bam_preseq -// -// output: -// file "*.ccurve.txt" into ch_mlib_preseq_mqc -// -// script: -// prefix = "${name}.mLb.mkD" -// """ -// preseq lc_extrap -v -output ${prefix}.ccurve.txt -bam ${bam[0]} -// """ -// } -// -// /* -// * STEP 5.2 Picard CollectMultipleMetrics after merging libraries and filtering -// */ -// process MergedLibMetrics { -// tag "$name" -// label 'process_medium' -// publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', -// saveAs: { filename -> -// if (filename.endsWith("_metrics")) "picard_metrics/$filename" -// else if (filename.endsWith(".pdf")) "picard_metrics/pdf/$filename" -// else null -// } -// -// when: -// !params.skip_picard_metrics -// -// input: -// set val(name), file(bam) from ch_mlib_rm_orphan_bam_metrics -// file fasta from ch_fasta -// -// output: -// file "*_metrics" into ch_mlib_collectmetrics_mqc -// file "*.pdf" into ch_mlib_collectmetrics_pdf -// -// script: -// prefix = "${name}.mLb.clN" -// def avail_mem = 3 -// if (!task.memory) { -// log.info "[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." -// } else { -// avail_mem = task.memory.toGiga() -// } -// """ -// picard -Xmx${avail_mem}g CollectMultipleMetrics \\ -// INPUT=${bam[0]} \\ -// OUTPUT=${prefix}.CollectMultipleMetrics \\ -// REFERENCE_SEQUENCE=$fasta \\ -// VALIDATION_STRINGENCY=LENIENT \\ -// TMP_DIR=tmp -// """ -// } -// -// /* -// * STEP 5.3 Read depth normalised bigWig -// */ -// process MergedLibBigWig { -// tag "$name" -// label 'process_medium' -// publishDir "${params.outdir}/bwa/mergedLibrary/bigwig", mode: 'copy', -// saveAs: { filename -> -// if (filename.endsWith("scale_factor.txt")) "scale/$filename" -// else if (filename.endsWith(".bigWig")) "$filename" -// else null -// } -// -// input: -// set val(name), file(bam), file(flagstat) from ch_mlib_rm_orphan_bam_bigwig.join(ch_mlib_rm_orphan_flagstat_bigwig, by: [0]) -// file sizes from ch_genome_sizes_mlib_bigwig.collect() -// -// output: -// set val(name), file("*.bigWig") into ch_mlib_bigwig_plotprofile -// file "*scale_factor.txt" into ch_mlib_bigwig_scale -// file "*igv.txt" into ch_mlib_bigwig_igv -// -// script: -// prefix = "${name}.mLb.clN" -// pe_fragment = params.single_end ? "" : "-pc" -// extend = (params.single_end && params.fragment_size > 0) ? "-fs ${params.fragment_size}" : '' -// """ -// SCALE_FACTOR=\$(grep 'mapped (' $flagstat | awk '{print 1000000/\$1}') -// echo \$SCALE_FACTOR > ${prefix}.scale_factor.txt -// genomeCoverageBed -ibam ${bam[0]} -bg -scale \$SCALE_FACTOR $pe_fragment $extend | sort -T '.' -k1,1 -k2,2n > ${prefix}.bedGraph -// -// bedGraphToBigWig ${prefix}.bedGraph $sizes ${prefix}.bigWig -// -// find * -type f -name "*.bigWig" -exec echo -e "bwa/mergedLibrary/bigwig/"{}"\\t0,0,178" \\; > ${prefix}.bigWig.igv.txt -// """ -// } -// -// /* -// * STEP 5.4 generate gene body coverage plot with deepTools -// */ -// process MergedLibPlotProfile { -// tag "$name" -// label 'process_high' -// publishDir "${params.outdir}/bwa/mergedLibrary/deepTools/plotProfile", mode: 'copy' -// -// when: -// !params.skip_plot_profile -// -// input: -// set val(name), file(bigwig) from ch_mlib_bigwig_plotprofile -// file bed from ch_gene_bed -// -// output: -// file '*.{gz,pdf}' into ch_mlib_plotprofile_results -// file '*.plotProfile.tab' into ch_mlib_plotprofile_mqc -// -// script: -// prefix = "${name}.mLb.clN" -// """ -// computeMatrix scale-regions \\ -// --regionsFileName $bed \\ -// --scoreFileName $bigwig \\ -// --outFileName ${prefix}.computeMatrix.mat.gz \\ -// --outFileNameMatrix ${prefix}.computeMatrix.vals.mat.gz \\ -// --regionBodyLength 1000 \\ -// --beforeRegionStartLength 3000 \\ -// --afterRegionStartLength 3000 \\ -// --skipZeros \\ -// --smartLabels \\ -// --numberOfProcessors $task.cpus -// -// plotProfile --matrixFile ${prefix}.computeMatrix.mat.gz \\ -// --outFileName ${prefix}.plotProfile.pdf \\ -// --outFileNameData ${prefix}.plotProfile.tab -// """ -// } -// -// /* -// * STEP 5.5 deepTools plotFingerprint -// */ -// process MergedLibPlotFingerprint { -// tag "$name" -// label 'process_high' -// publishDir "${params.outdir}/bwa/mergedLibrary/deepTools/plotFingerprint", mode: 'copy' -// -// when: -// !params.skip_plot_fingerprint -// -// input: -// set val(name), file(bam) from ch_mlib_rm_orphan_bam_plotfingerprint -// -// output: -// file '*.{txt,pdf}' into ch_mlib_plotfingerprint_results -// file '*.raw.txt' into ch_mlib_plotfingerprint_mqc -// -// script: -// prefix = "${name}.mLb.clN" -// extend = (params.single_end && params.fragment_size > 0) ? "--extendReads ${params.fragment_size}" : '' -// """ -// plotFingerprint \\ -// --bamfiles ${bam[0]} \\ -// --plotFile ${prefix}.plotFingerprint.pdf \\ -// $extend \\ -// --labels $prefix \\ -// --outRawCounts ${prefix}.plotFingerprint.raw.txt \\ -// --outQualityMetrics ${prefix}.plotFingerprint.qcmetrics.txt \\ -// --skipZeros \\ -// --numberOfProcessors $task.cpus \\ -// --numberOfSamples $params.fingerprint_bins -// """ -// } -// -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// /* -- -- */ -// /* -- MERGE LIBRARY PEAK ANALYSIS -- */ -// /* -- -- */ -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// -// /* -// * STEP 6.1 Call peaks with MACS2 and calculate FRiP score -// */ -// process MergedLibMACSCallPeak { -// tag "$name" -// label 'process_medium' -// publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}", mode: 'copy', -// saveAs: { filename -> -// if (filename.endsWith(".tsv")) "qc/$filename" -// else if (filename.endsWith(".igv.txt")) null -// else filename -// } -// -// when: -// params.macs_gsize -// -// input: -// set val(name), file(bam), file(flagstat) from ch_mlib_rm_orphan_bam_macs.join(ch_mlib_rm_orphan_flagstat_macs, by: [0]) -// file mlib_peak_count_header from ch_mlib_peak_count_header -// file mlib_frip_score_header from ch_mlib_frip_score_header -// -// output: -// set val(name), file("*.{bed,xls,gappedPeak,bdg}") into ch_mlib_macs_output -// set val(name), file("*$PEAK_TYPE") into ch_mlib_macs_homer, -// ch_mlib_macs_qc, -// ch_mlib_macs_consensus, -// ch_mlib_macs_ataqv -// file "*igv.txt" into ch_mlib_macs_igv -// file "*_mqc.tsv" into ch_mlib_macs_mqc -// -// script: -// prefix = "${name}.mLb.clN" -// broad = params.narrow_peak ? '' : "--broad --broad-cutoff ${params.broad_cutoff}" -// format = params.single_end ? "BAM" : "BAMPE" -// pileup = params.save_macs_pileup ? "-B --SPMR" : "" -// """ -// macs2 callpeak \\ -// -t ${bam[0]} \\ -// $broad \\ -// -f $format \\ -// -g $params.macs_gsize \\ -// -n $prefix \\ -// $pileup \\ -// --keep-dup all \\ -// --nomodel -// -// cat ${prefix}_peaks.${PEAK_TYPE} | wc -l | awk -v OFS='\t' '{ print "${name}", \$1 }' | cat $mlib_peak_count_header - > ${prefix}_peaks.count_mqc.tsv -// -// READS_IN_PEAKS=\$(intersectBed -a ${bam[0]} -b ${prefix}_peaks.${PEAK_TYPE} -bed -c -f 0.20 | awk -F '\t' '{sum += \$NF} END {print sum}') -// grep 'mapped (' $flagstat | awk -v a="\$READS_IN_PEAKS" -v OFS='\t' '{print "${name}", a/\$1}' | cat $mlib_frip_score_header - > ${prefix}_peaks.FRiP_mqc.tsv -// -// find * -type f -name "*.${PEAK_TYPE}" -exec echo -e "bwa/mergedLibrary/macs/${PEAK_TYPE}/"{}"\\t0,0,178" \\; > ${prefix}_peaks.igv.txt -// """ -// } -// -// /* -// * STEP 6.2 Annotate peaks with HOMER -// */ -// process MergedLibAnnotatePeaks { -// tag "$name" -// label "process_medium" -// publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}", mode: 'copy' -// -// when: -// params.macs_gsize -// -// input: -// set val(name), file(peak) from ch_mlib_macs_homer -// file fasta from ch_fasta -// file gtf from ch_gtf -// -// output: -// file "*.txt" into ch_mlib_macs_annotate -// -// script: -// prefix = "${name}.mLb.clN" -// """ -// annotatePeaks.pl \\ -// $peak \\ -// $fasta \\ -// -gid \\ -// -gtf $gtf \\ -// -cpu $task.cpus \\ -// > ${prefix}_peaks.annotatePeaks.txt -// """ -// } -// -// /* -// * STEP 6.3 Aggregated QC plots for peaks, FRiP and peak-to-gene annotation -// */ -// process MergedLibPeakQC { -// label "process_medium" -// publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/qc", mode: 'copy' -// -// when: -// params.macs_gsize -// -// input: -// file peaks from ch_mlib_macs_qc.collect{ it[1] } -// file annos from ch_mlib_macs_annotate.collect() -// file mlib_peak_annotation_header from ch_mlib_peak_annotation_header -// -// output: -// file "*.{txt,pdf}" into ch_mlib_peak_qc -// file "*.tsv" into ch_mlib_peak_qc_mqc -// -// script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ -// suffix = 'mLb.clN' -// """ -// plot_macs_qc.r \\ -// -i ${peaks.join(',')} \\ -// -s ${peaks.join(',').replaceAll(".${suffix}_peaks.${PEAK_TYPE}","")} \\ -// -o ./ \\ -// -p macs_peak.${suffix} -// -// plot_homer_annotatepeaks.r \\ -// -i ${annos.join(',')} \\ -// -s ${annos.join(',').replaceAll(".${suffix}_peaks.annotatePeaks.txt","")} \\ -// -o ./ \\ -// -p macs_annotatePeaks.${suffix} -// -// cat $mlib_peak_annotation_header macs_annotatePeaks.${suffix}.summary.txt > macs_annotatePeaks.${suffix}.summary_mqc.tsv -// """ -// } -// -// /* -// * STEP 6.4 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection -// */ -// process MergedLibConsensusPeakSet { -// label 'process_long' -// publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: 'copy', -// saveAs: { filename -> -// if (filename.endsWith(".igv.txt")) null -// else filename -// } -// -// when: -// params.macs_gsize && (replicatesExist || multipleGroups) && !params.skip_consensus_peaks -// -// input: -// file peaks from ch_mlib_macs_consensus.collect{ it[1] } -// -// output: -// file "*.bed" into ch_mlib_macs_consensus_bed -// file "*.saf" into ch_mlib_macs_consensus_saf -// file "*.boolean.txt" into ch_mlib_macs_consensus_bool -// file "*.intersect.{txt,plot.pdf}" into ch_mlib_macs_consensus_intersect -// file "*igv.txt" into ch_mlib_macs_consensus_igv -// -// script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ -// suffix = 'mLb.clN' -// prefix = "consensus_peaks.${suffix}" -// mergecols = params.narrow_peak ? (2..10).join(',') : (2..9).join(',') -// collapsecols = params.narrow_peak ? (["collapse"]*9).join(',') : (["collapse"]*8).join(',') -// expandparam = params.narrow_peak ? "--is_narrow_peak" : "" -// """ -// sort -T '.' -k1,1 -k2,2n ${peaks.collect{it.toString()}.sort().join(' ')} \\ -// | mergeBed -c $mergecols -o $collapsecols > ${prefix}.txt -// -// macs2_merged_expand.py \\ -// ${prefix}.txt \\ -// ${peaks.collect{it.toString()}.sort().join(',').replaceAll("_peaks.${PEAK_TYPE}","")} \\ -// ${prefix}.boolean.txt \\ -// --min_replicates $params.min_reps_consensus \\ -// $expandparam -// -// awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$1, \$2, \$3, \$4, "0", "+" }' ${prefix}.boolean.txt > ${prefix}.bed -// -// echo -e "GeneID\tChr\tStart\tEnd\tStrand" > ${prefix}.saf -// awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$4, \$1, \$2, \$3, "+" }' ${prefix}.boolean.txt >> ${prefix}.saf -// -// sed -i 's/.${suffix}//g' ${prefix}.boolean.intersect.txt -// plot_peak_intersect.r -i ${prefix}.boolean.intersect.txt -o ${prefix}.boolean.intersect.plot.pdf -// -// find * -type f -name "${prefix}.bed" -exec echo -e "bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus/"{}"\\t0,0,0" \\; > ${prefix}.bed.igv.txt -// """ -// } -// -// /* -// * STEP 6.5 Annotate consensus peaks with HOMER, and add annotation to boolean output file -// */ -// process MergedLibConsensusPeakSetAnnotate { -// label "process_medium" -// publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: 'copy' -// -// when: -// params.macs_gsize && (replicatesExist || multipleGroups) && !params.skip_consensus_peaks -// -// input: -// file bed from ch_mlib_macs_consensus_bed -// file bool from ch_mlib_macs_consensus_bool -// file fasta from ch_fasta -// file gtf from ch_gtf -// -// output: -// file "*.annotatePeaks.txt" into ch_mlib_macs_consensus_annotate -// -// script: -// prefix = "consensus_peaks.mLb.clN" -// """ -// annotatePeaks.pl \\ -// $bed \\ -// $fasta \\ -// -gid \\ -// -gtf $gtf \\ -// -cpu $task.cpus \\ -// > ${prefix}.annotatePeaks.txt -// -// cut -f2- ${prefix}.annotatePeaks.txt | awk 'NR==1; NR > 1 {print \$0 | "sort -T '.' -k1,1 -k2,2n"}' | cut -f6- > tmp.txt -// paste $bool tmp.txt > ${prefix}.boolean.annotatePeaks.txt -// """ -// } -// -// /* -// * STEP 6.6 Count reads in consensus peaks with featureCounts and perform differential analysis with DESeq2 -// */ -// process MergedLibConsensusPeakSetDESeq { -// label 'process_medium' -// publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus/deseq2", mode: 'copy', -// saveAs: { filename -> -// if (filename.endsWith(".igv.txt")) null -// else filename -// } -// -// when: -// params.macs_gsize && replicatesExist && multipleGroups && !params.skip_consensus_peaks -// -// input: -// file bams from ch_mlib_name_bam_mlib_counts.collect{ it[1] } -// file saf from ch_mlib_macs_consensus_saf.collect() -// file mlib_deseq2_pca_header from ch_mlib_deseq2_pca_header -// file mlib_deseq2_clustering_header from ch_mlib_deseq2_clustering_header -// -// output: -// file "*featureCounts.txt" into ch_mlib_macs_consensus_counts -// file "*featureCounts.txt.summary" into ch_mlib_macs_consensus_counts_mqc -// file "*.{RData,results.txt,pdf,log}" into ch_mlib_macs_consensus_deseq_results -// file "sizeFactors" into ch_mlib_macs_consensus_deseq_factors -// file "*vs*/*.{pdf,txt}" into ch_mlib_macs_consensus_deseq_comp_results -// file "*vs*/*.bed" into ch_mlib_macs_consensus_deseq_comp_bed -// file "*igv.txt" into ch_mlib_macs_consensus_deseq_comp_igv -// file "*.tsv" into ch_mlib_macs_consensus_deseq_mqc -// -// script: -// prefix = "consensus_peaks.mLb.clN" -// bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() -// bam_ext = params.single_end ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" -// pe_params = params.single_end ? '' : "-p --donotsort" -// """ -// featureCounts \\ -// -F SAF \\ -// -O \\ -// --fracOverlap 0.2 \\ -// -T $task.cpus \\ -// $pe_params \\ -// -a $saf \\ -// -o ${prefix}.featureCounts.txt \\ -// ${bam_files.join(' ')} -// -// featurecounts_deseq2.r -i ${prefix}.featureCounts.txt -b '$bam_ext' -o ./ -p $prefix -s .mLb -// -// cat $mlib_deseq2_pca_header ${prefix}.pca.vals.txt > ${prefix}.pca.vals_mqc.tsv -// cat $mlib_deseq2_clustering_header ${prefix}.sample.dists.txt > ${prefix}.sample.dists_mqc.tsv -// -// find * -type f -name "*.FDR0.05.results.bed" -exec echo -e "bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus/deseq2/"{}"\\t255,0,0" \\; > ${prefix}.igv.txt -// """ -// } -// -// /* -// * STEP 6.7 Run ataqv on BAM file and corresponding peaks -// */ -// process MergedLibAtaqv { -// tag "$name" -// label 'process_medium' -// publishDir "${params.outdir}/bwa/mergedLibrary/ataqv/${PEAK_TYPE}", mode: 'copy' -// -// when: -// !params.skip_ataqv -// -// input: -// set val(name), file(bam), file(peak) from ch_mlib_bam_ataqv.join(ch_mlib_macs_ataqv, by: [0]) -// file autosomes from ch_genome_autosomes.collect() -// file tss_bed from ch_tss_bed -// -// output: -// file "*.json" into ch_mlib_ataqv -// -// script: -// peak_param = params.macs_gsize ? "--peak-file ${peak}" : "" -// mito_param = params.mito_name ? "--mitochondrial-reference-name ${params.mito_name}" : "" -// """ -// ataqv \\ -// --threads $task.cpus \\ -// $peak_param \\ -// --tss-file $tss_bed \\ -// --metrics-file ${name}.ataqv.json \\ -// --name $name \\ -// --ignore-read-groups \\ -// --autosomal-reference-file $autosomes \\ -// $mito_param \\ -// NA \\ -// ${bam[0]} -// """ -// } -// -// /* -// * STEP 6.8 run ataqv mkarv on all JSON files to render web app -// */ -// process MergedLibAtaqvMkarv { -// label 'process_medium' -// publishDir "${params.outdir}/bwa/mergedLibrary/ataqv/${PEAK_TYPE}", mode: 'copy' -// -// when: -// !params.skip_ataqv -// -// input: -// file json from ch_mlib_ataqv.collect() -// -// output: -// file "html" into ch_mlib_ataqv_mkarv -// -// script: -// """ -// mkarv \\ -// --concurrency $task.cpus \\ -// --force \\ -// ./html/ \\ -// ${json.join(' ')} -// """ -// } -// -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// /* -- -- */ -// /* -- MERGE REPLICATE BAM -- */ -// /* -- -- */ -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// -// /* -// * STEP 7 Merge library BAM files across all replicates -// */ -// ch_mlib_rm_orphan_bam_mrep -// .map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] } -// .groupTuple(by: [0]) -// .map { it -> [ it[0], it[1].flatten() ] } -// .set { ch_mlib_rm_orphan_bam_mrep } -// -// process MergedRepBAM { -// tag "$name" -// label 'process_medium' -// publishDir "${params.outdir}/bwa/mergedReplicate", mode: 'copy', -// saveAs: { filename -> -// if (filename.endsWith(".flagstat")) "samtools_stats/$filename" -// else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" -// else if (filename.endsWith(".stats")) "samtools_stats/$filename" -// else if (filename.endsWith(".metrics.txt")) "picard_metrics/$filename" -// else filename -// } -// -// input: -// set val(name), file(bams) from ch_mlib_rm_orphan_bam_mrep -// -// output: -// set val(name), file("*${prefix}.sorted.{bam,bam.bai}") into ch_mrep_bam_bigwig, -// ch_mrep_bam_macs -// set val(name), file("*.flagstat") into ch_mrep_bam_flagstat_bigwig, -// ch_mrep_bam_flagstat_macs, -// ch_mrep_bam_flagstat_mqc -// file "*.{idxstats,stats}" into ch_mrep_bam_stats_mqc -// file "*.txt" into ch_mrep_bam_metrics_mqc -// -// when: -// !params.skip_merge_replicates && replicatesExist -// -// script: -// prefix = "${name}.mRp.clN" -// bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() -// def avail_mem = 3 -// if (!task.memory) { -// log.info "[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." -// } else { -// avail_mem = task.memory.toGiga() -// } -// if (bam_files.size() > 1) { -// """ -// picard -Xmx${avail_mem}g MergeSamFiles \\ -// ${'INPUT='+bam_files.join(' INPUT=')} \\ -// OUTPUT=${name}.sorted.bam \\ -// SORT_ORDER=coordinate \\ -// VALIDATION_STRINGENCY=LENIENT \\ -// TMP_DIR=tmp -// samtools index ${name}.sorted.bam -// -// picard -Xmx${avail_mem}g MarkDuplicates \\ -// INPUT=${name}.sorted.bam \\ -// OUTPUT=${prefix}.sorted.bam \\ -// ASSUME_SORTED=true \\ -// REMOVE_DUPLICATES=true \\ -// METRICS_FILE=${prefix}.MarkDuplicates.metrics.txt \\ -// VALIDATION_STRINGENCY=LENIENT \\ -// TMP_DIR=tmp -// -// samtools index ${prefix}.sorted.bam -// samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat -// samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats -// samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats -// """ -// } else { -// """ -// ln -s ${bams[0]} ${prefix}.sorted.bam -// ln -s ${bams[1]} ${prefix}.sorted.bam.bai -// touch ${prefix}.MarkDuplicates.metrics.txt -// samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat -// samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats -// samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats -// """ -// } -// } -// -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// /* -- -- */ -// /* -- MERGE REPLICATE BAM POST-ANALYSIS -- */ -// /* -- -- */ -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// -// /* -// * STEP 8.1 Read depth normalised bigWig -// */ -// process MergedRepBigWig { -// tag "$name" -// label 'process_medium' -// publishDir "${params.outdir}/bwa/mergedReplicate/bigwig", mode: 'copy', -// saveAs: { filename -> -// if (filename.endsWith("scale_factor.txt")) "scale/$filename" -// else if (filename.endsWith(".bigWig")) "$filename" -// else null -// } -// -// when: -// !params.skip_merge_replicates && replicatesExist -// -// input: -// set val(name), file(bam), file(flagstat) from ch_mrep_bam_bigwig.join(ch_mrep_bam_flagstat_bigwig, by: [0]) -// file sizes from ch_genome_sizes_mrep_bigwig.collect() -// -// output: -// set val(name), file("*.bigWig") into ch_mrep_bigwig -// file "*scale_factor.txt" into ch_mrep_bigwig_scale -// file "*igv.txt" into ch_mrep_bigwig_igv -// -// script: -// prefix = "${name}.mRp.clN" -// pe_fragment = params.single_end ? "" : "-pc" -// extend = (params.single_end && params.fragment_size > 0) ? "-fs ${params.fragment_size}" : '' -// """ -// SCALE_FACTOR=\$(grep 'mapped (' $flagstat | awk '{print 1000000/\$1}') -// echo \$SCALE_FACTOR > ${prefix}.scale_factor.txt -// genomeCoverageBed -ibam ${bam[0]} -bg -scale \$SCALE_FACTOR $pe_fragment $extend | sort -T '.' -k1,1 -k2,2n > ${prefix}.bedGraph -// -// bedGraphToBigWig ${prefix}.bedGraph $sizes ${prefix}.bigWig -// -// find * -type f -name "*.bigWig" -exec echo -e "bwa/mergedReplicate/bigwig/"{}"\\t0,0,178" \\; > ${prefix}.bigWig.igv.txt -// """ -// } -// -// /* -// * STEP 8.2 Call peaks with MACS2 and calculate FRiP score -// */ -// process MergedRepMACSCallPeak { -// tag "$name" -// label 'process_medium' -// publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}", mode: 'copy', -// saveAs: { filename -> -// if (filename.endsWith(".tsv")) "qc/$filename" -// else if (filename.endsWith(".igv.txt")) null -// else filename -// } -// -// when: -// !params.skip_merge_replicates && replicatesExist && params.macs_gsize -// -// input: -// set val(name), file(bam), file(flagstat) from ch_mrep_bam_macs.join(ch_mrep_bam_flagstat_macs, by: [0]) -// file mrep_peak_count_header from ch_mrep_peak_count_header -// file mrep_frip_score_header from ch_mrep_frip_score_header -// -// output: -// set val(name), file("*.{bed,xls,gappedPeak,bdg}") into ch_mrep_macs_output -// set val(name), file("*$PEAK_TYPE") into ch_mrep_macs_homer, -// ch_mrep_macs_qc, -// ch_mrep_macs_consensus -// file "*igv.txt" into ch_mrep_macs_igv -// file "*_mqc.tsv" into ch_mrep_macs_mqc -// -// script: -// prefix = "${name}.mRp.clN" -// broad = params.narrow_peak ? '' : "--broad --broad-cutoff ${params.broad_cutoff}" -// format = params.single_end ? "BAM" : "BAMPE" -// pileup = params.save_macs_pileup ? "-B --SPMR" : "" -// """ -// macs2 callpeak \\ -// -t ${bam[0]} \\ -// $broad \\ -// -f $format \\ -// -g $params.macs_gsize \\ -// -n $prefix \\ -// $pileup \\ -// --keep-dup all \\ -// --nomodel -// -// cat ${prefix}_peaks.${PEAK_TYPE} | wc -l | awk -v OFS='\t' '{ print "${name}", \$1 }' | cat $mrep_peak_count_header - > ${prefix}_peaks.count_mqc.tsv -// -// READS_IN_PEAKS=\$(intersectBed -a ${bam[0]} -b ${prefix}_peaks.${PEAK_TYPE} -bed -c -f 0.20 | awk -F '\t' '{sum += \$NF} END {print sum}') -// grep 'mapped (' $flagstat | awk -v a="\$READS_IN_PEAKS" -v OFS='\t' '{print "${name}", a/\$1}' | cat $mrep_frip_score_header - > ${prefix}_peaks.FRiP_mqc.tsv -// -// find * -type f -name "*.${PEAK_TYPE}" -exec echo -e "bwa/mergedReplicate/macs/${PEAK_TYPE}/"{}"\\t0,0,178" \\; > ${prefix}_peaks.igv.txt -// """ -// } -// -// /* -// * STEP 8.3 Annotate peaks with HOMER -// */ -// process MergedRepAnnotatePeaks { -// tag "$name" -// label "process_medium" -// publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}", mode: 'copy' -// -// when: -// !params.skip_merge_replicates && replicatesExist && params.macs_gsize -// -// input: -// set val(name), file(peak) from ch_mrep_macs_homer -// file fasta from ch_fasta -// file gtf from ch_gtf -// -// output: -// file "*.txt" into ch_mrep_macs_annotate -// -// script: -// prefix = "${name}.mRp.clN" -// """ -// annotatePeaks.pl \\ -// $peak \\ -// $fasta \\ -// -gid \\ -// -gtf $gtf \\ -// -cpu $task.cpus \\ -// > ${prefix}_peaks.annotatePeaks.txt -// """ -// } -// -// /* -// * STEP 8.4 Aggregated QC plots for peaks, FRiP and peak-to-gene annotation -// */ -// process MergedRepPeakQC { -// label "process_medium" -// publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/qc", mode: 'copy' -// -// when: -// !params.skip_merge_replicates && replicatesExist && params.macs_gsize -// -// input: -// file peaks from ch_mrep_macs_qc.collect{ it[1] } -// file annos from ch_mrep_macs_annotate.collect() -// file mrep_peak_annotation_header from ch_mrep_peak_annotation_header -// -// output: -// file "*.{txt,pdf}" into ch_mrep_peak_qc -// file "*.tsv" into ch_mrep_peak_qc_mqc -// -// script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ -// suffix = 'mRp.clN' -// """ -// plot_macs_qc.r \\ -// -i ${peaks.join(',')} \\ -// -s ${peaks.join(',').replaceAll(".${suffix}_peaks.${PEAK_TYPE}","")} \\ -// -o ./ \\ -// -p macs_peak.${suffix} -// -// plot_homer_annotatepeaks.r \\ -// -i ${annos.join(',')} \\ -// -s ${annos.join(',').replaceAll(".${suffix}_peaks.annotatePeaks.txt","")} \\ -// -o ./ \\ -// -p macs_annotatePeaks.${suffix} -// -// cat $mrep_peak_annotation_header macs_annotatePeaks.${suffix}.summary.txt > macs_annotatePeaks.${suffix}.summary_mqc.tsv -// """ -// } -// -// /* -// * STEP 8.5 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection -// */ -// process MergedRepConsensusPeakSet { -// label 'process_long' -// publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: 'copy', -// saveAs: { filename -> -// if (filename.endsWith(".igv.txt")) null -// else filename -// } -// -// when: -// !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks -// -// input: -// file peaks from ch_mrep_macs_consensus.collect{ it[1] } -// -// output: -// file "*.bed" into ch_mrep_macs_consensus_bed -// file "*.saf" into ch_mrep_macs_consensus_saf -// file "*.boolean.txt" into ch_mrep_macs_consensus_bool -// file "*.intersect.{txt,plot.pdf}" into ch_mrep_macs_consensus_intersect -// file "*igv.txt" into ch_mrep_macs_consensus_igv -// -// script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ -// suffix = 'mRp.clN' -// prefix = "consensus_peaks.${suffix}" -// mergecols = params.narrow_peak ? (2..10).join(',') : (2..9).join(',') -// collapsecols = params.narrow_peak ? (["collapse"]*9).join(',') : (["collapse"]*8).join(',') -// expandparam = params.narrow_peak ? "--is_narrow_peak" : "" -// """ -// sort -T '.' -k1,1 -k2,2n ${peaks.collect{it.toString()}.sort().join(' ')} \\ -// | mergeBed -c $mergecols -o $collapsecols > ${prefix}.txt -// -// macs2_merged_expand.py \\ -// ${prefix}.txt \\ -// ${peaks.collect{it.toString()}.sort().join(',').replaceAll("_peaks.${PEAK_TYPE}","")} \\ -// ${prefix}.boolean.txt \\ -// $expandparam -// -// awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$1, \$2, \$3, \$4, "0", "+" }' ${prefix}.boolean.txt > ${prefix}.bed -// -// echo -e "GeneID\tChr\tStart\tEnd\tStrand" > ${prefix}.saf -// awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$4, \$1, \$2, \$3, "+" }' ${prefix}.boolean.txt >> ${prefix}.saf -// -// sed -i 's/.${suffix}//g' ${prefix}.boolean.intersect.txt -// plot_peak_intersect.r -i ${prefix}.boolean.intersect.txt -o ${prefix}.boolean.intersect.plot.pdf -// -// find * -type f -name "${prefix}.bed" -exec echo -e "bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus/"{}"\\t0,0,0" \\; > ${prefix}.bed.igv.txt -// """ -// } -// -// /* -// * STEP 8.6 Annotate consensus peaks with HOMER, and add annotation to boolean output file -// */ -// process MergedRepConsensusPeakSetAnnotate { -// label "process_medium" -// publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: 'copy' -// -// when: -// !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks -// -// input: -// file bed from ch_mrep_macs_consensus_bed -// file bool from ch_mrep_macs_consensus_bool -// file fasta from ch_fasta -// file gtf from ch_gtf -// -// output: -// file "*.annotatePeaks.txt" into ch_mrep_macs_consensus_annotate -// -// script: -// prefix = "consensus_peaks.mRp.clN" -// """ -// annotatePeaks.pl \\ -// $bed \\ -// $fasta \\ -// -gid \\ -// -gtf $gtf \\ -// -cpu $task.cpus \\ -// > ${prefix}.annotatePeaks.txt -// -// cut -f2- ${prefix}.annotatePeaks.txt | awk 'NR==1; NR > 1 {print \$0 | "sort -T '.' -k1,1 -k2,2n"}' | cut -f6- > tmp.txt -// paste $bool tmp.txt > ${prefix}.boolean.annotatePeaks.txt -// """ -// } -// -// /* -// * STEP 8.7 Count reads in consensus peaks with featureCounts and perform differential analysis with DESeq2 -// */ -// process MergedRepConsensusPeakSetDESeq { -// label 'process_medium' -// publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus/deseq2", mode: 'copy', -// saveAs: { filename -> -// if (filename.endsWith(".igv.txt")) null -// else filename -// } -// -// when: -// !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks -// -// input: -// file bams from ch_mlib_name_bam_mrep_counts.collect{ it[1] } -// file saf from ch_mrep_macs_consensus_saf.collect() -// file mrep_deseq2_pca_header from ch_mrep_deseq2_pca_header -// file mrep_deseq2_clustering_header from ch_mrep_deseq2_clustering_header -// -// output: -// file "*featureCounts.txt" into ch_mrep_macs_consensus_counts -// file "*featureCounts.txt.summary" into ch_mrep_macs_consensus_counts_mqc -// file "*.{RData,results.txt,pdf,log}" into ch_mrep_macs_consensus_deseq_results -// file "sizeFactors" into ch_mrep_macs_consensus_deseq_factors -// file "*vs*/*.{pdf,txt}" into ch_mrep_macs_consensus_deseq_comp_results -// file "*vs*/*.bed" into ch_mrep_macs_consensus_deseq_comp_bed -// file "*igv.txt" into ch_mrep_macs_consensus_deseq_comp_igv -// file "*.tsv" into ch_mrep_macs_consensus_deseq_mqc -// -// script: -// prefix = "consensus_peaks.mRp.clN" -// bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() -// bam_ext = params.single_end ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" -// pe_params = params.single_end ? '' : "-p --donotsort" -// """ -// featureCounts \\ -// -F SAF \\ -// -O \\ -// --fracOverlap 0.2 \\ -// -T $task.cpus \\ -// $pe_params \\ -// -a $saf \\ -// -o ${prefix}.featureCounts.txt \\ -// ${bam_files.join(' ')} -// -// featurecounts_deseq2.r -i ${prefix}.featureCounts.txt -b '$bam_ext' -o ./ -p $prefix -s .mLb -// -// cat $mrep_deseq2_pca_header ${prefix}.pca.vals.txt > ${prefix}.pca.vals_mqc.tsv -// cat $mrep_deseq2_clustering_header ${prefix}.sample.dists.txt > ${prefix}.sample.dists_mqc.tsv -// -// find * -type f -name "*.FDR0.05.results.bed" -exec echo -e "bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus/deseq2/"{}"\\t255,0,0" \\; > ${prefix}.igv.txt -// """ -// } -// -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// /* -- -- */ -// /* -- IGV -- */ -// /* -- -- */ -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// -// /* -// * STEP 9 - Create IGV session file -// */ -// process IGV { -// publishDir "${params.outdir}/igv/${PEAK_TYPE}", mode: 'copy' -// -// when: -// !params.skip_igv -// -// input: -// file fasta from ch_fasta -// -// file bigwigs from ch_mlib_bigwig_igv.collect().ifEmpty([]) -// file peaks from ch_mlib_macs_igv.collect().ifEmpty([]) -// file consensus_peaks from ch_mlib_macs_consensus_igv.collect().ifEmpty([]) -// file differential_peaks from ch_mlib_macs_consensus_deseq_comp_igv.collect().ifEmpty([]) -// -// file rbigwigs from ch_mrep_bigwig_igv.collect().ifEmpty([]) -// file rpeaks from ch_mrep_macs_igv.collect().ifEmpty([]) -// file rconsensus_peaks from ch_mrep_macs_consensus_igv.collect().ifEmpty([]) -// file rdifferential_peaks from ch_mrep_macs_consensus_deseq_comp_igv.collect().ifEmpty([]) -// -// output: -// file "*.{txt,xml}" into ch_igv_session -// -// script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ -// """ -// cat *.txt > igv_files.txt -// igv_files_to_session.py igv_session.xml igv_files.txt ../../reference_genome/${fasta.getName()} --path_prefix '../../' -// """ -// } -// -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// /* -- -- */ -// /* -- MULTIQC -- */ -// /* -- -- */ -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// -// /* -// * Parse software version numbers -// */ -// process get_software_versions { -// publishDir "${params.outdir}/pipeline_info", mode: 'copy', -// saveAs: { filename -> -// if (filename.indexOf(".csv") > 0) filename -// else null -// } -// -// output: -// file 'software_versions_mqc.yaml' into ch_software_versions_mqc -// file "software_versions.csv" -// -// script: -// """ -// echo $workflow.manifest.version > v_pipeline.txt -// echo $workflow.nextflow.version > v_nextflow.txt -// fastqc --version > v_fastqc.txt -// trim_galore --version > v_trim_galore.txt -// echo \$(bwa 2>&1) > v_bwa.txt -// samtools --version > v_samtools.txt -// bedtools --version > v_bedtools.txt -// echo \$(bamtools --version 2>&1) > v_bamtools.txt -// echo \$(plotFingerprint --version 2>&1) > v_deeptools.txt || true -// picard MarkDuplicates --version &> v_picard.txt || true -// echo \$(R --version 2>&1) > v_R.txt -// python -c "import pysam; print(pysam.__version__)" > v_pysam.txt -// echo \$(macs2 --version 2>&1) > v_macs2.txt -// touch v_homer.txt -// echo \$(ataqv --version 2>&1) > v_ataqv.txt -// echo \$(featureCounts -v 2>&1) > v_featurecounts.txt -// preseq &> v_preseq.txt -// multiqc --version > v_multiqc.txt -// scrape_software_versions.py &> software_versions_mqc.yaml -// """ -// } -// -// def create_workflow_summary(summary) { -// -// def yaml_file = workDir.resolve('workflow_summary_mqc.yaml') -// yaml_file.text = """ -// id: 'nf-core-atacseq-summary' -// description: " - this information is collected when the pipeline is started." -// section_name: 'nf-core/atacseq Workflow Summary' -// section_href: 'https://github.com/nf-core/atacseq' -// plot_type: 'html' -// data: | -//
-// ${summary.collect { k,v -> "
$k
${v ?: 'N/A'}
" }.join("\n")} -//
-// """.stripIndent() -// -// return yaml_file -// } -// -// /* -// * STEP 10 - MultiQC -// */ -// process MultiQC { -// publishDir "${params.outdir}/multiqc/${PEAK_TYPE}", mode: 'copy' -// -// when: -// !params.skip_multiqc -// -// input: -// file multiqc_config from ch_multiqc_config -// -// file ('software_versions/*') from ch_software_versions_mqc.collect() -// file ('workflow_summary/*') from create_workflow_summary(summary) -// -// file ('fastqc/*') from ch_fastqc_reports_mqc.collect().ifEmpty([]) -// file ('trimgalore/*') from ch_trimgalore_results_mqc.collect().ifEmpty([]) -// file ('trimgalore/fastqc/*') from ch_trimgalore_fastqc_reports_mqc.collect().ifEmpty([]) -// -// file ('alignment/library/*') from ch_sort_bam_flagstat_mqc.collect() -// -// file ('alignment/mergedLibrary/*') from ch_mlib_bam_stats_mqc.collect() -// file ('alignment/mergedLibrary/*') from ch_mlib_rm_orphan_flagstat_mqc.collect{it[1]} -// file ('alignment/mergedLibrary/*') from ch_mlib_rm_orphan_stats_mqc.collect() -// file ('alignment/mergedLibrary/picard_metrics/*') from ch_mlib_bam_metrics_mqc.collect() -// file ('alignment/mergedLibrary/picard_metrics/*') from ch_mlib_collectmetrics_mqc.collect() -// file ('macs/mergedLibrary/*') from ch_mlib_macs_mqc.collect().ifEmpty([]) -// file ('macs/mergedLibrary/*') from ch_mlib_peak_qc_mqc.collect().ifEmpty([]) -// file ('macs/mergedLibrary/consensus/*') from ch_mlib_macs_consensus_counts_mqc.collect().ifEmpty([]) -// file ('macs/mergedLibrary/consensus/*') from ch_mlib_macs_consensus_deseq_mqc.collect().ifEmpty([]) -// file ('preseq/*') from ch_mlib_preseq_mqc.collect().ifEmpty([]) -// file ('deeptools/*') from ch_mlib_plotprofile_mqc.collect().ifEmpty([]) -// file ('deeptools/*') from ch_mlib_plotfingerprint_mqc.collect().ifEmpty([]) -// -// file ('alignment/mergedReplicate/*') from ch_mrep_bam_flagstat_mqc.collect{it[1]}.ifEmpty([]) -// file ('alignment/mergedReplicate/*') from ch_mrep_bam_stats_mqc.collect().ifEmpty([]) -// file ('alignment/mergedReplicate/*') from ch_mrep_bam_metrics_mqc.collect().ifEmpty([]) -// file ('macs/mergedReplicate/*') from ch_mrep_macs_mqc.collect().ifEmpty([]) -// file ('macs/mergedReplicate/*') from ch_mrep_peak_qc_mqc.collect().ifEmpty([]) -// file ('macs/mergedReplicate/consensus/*') from ch_mrep_macs_consensus_counts_mqc.collect().ifEmpty([]) -// file ('macs/mergedReplicate/consensus/*') from ch_mrep_macs_consensus_deseq_mqc.collect().ifEmpty([]) -// -// output: -// file "*multiqc_report.html" into ch_multiqc_report -// file "*_data" -// file "multiqc_plots" -// -// script: -// rtitle = custom_runName ? "--title \"$custom_runName\"" : '' -// rfilename = custom_runName ? "--filename " + custom_runName.replaceAll('\\W','_').replaceAll('_+','_') + "_multiqc_report" : '' -// """ -// multiqc . -f $rtitle $rfilename --config $multiqc_config \\ -// -m custom_content -m fastqc -m cutadapt -m samtools -m picard -m preseq -m featureCounts -m deeptools -// """ -// } -// -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// /* -- -- */ -// /* -- REPORTS/DOCUMENTATION -- */ -// /* -- -- */ -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// -// /* -// * STEP 11 - Output description HTML -// */ -// process output_documentation { -// publishDir "${params.outdir}/Documentation", mode: 'copy' -// -// input: -// file output_docs from ch_output_docs -// -// output: -// file "results_description.html" -// -// script: -// """ -// markdown_to_html.r $output_docs results_description.html -// """ -// } -// -// /* -// * Completion e-mail notification -// */ -// workflow.onComplete { -// -// // Set up the e-mail variables -// def subject = "[nf-core/atacseq] Successful: $workflow.runName" -// if (!workflow.success) { -// subject = "[nf-core/atacseq] FAILED: $workflow.runName" -// } -// def email_fields = [:] -// email_fields['version'] = workflow.manifest.version -// email_fields['runName'] = custom_runName ?: workflow.runName -// email_fields['success'] = workflow.success -// email_fields['dateComplete'] = workflow.complete -// email_fields['duration'] = workflow.duration -// email_fields['exitStatus'] = workflow.exitStatus -// email_fields['errorMessage'] = (workflow.errorMessage ?: 'None') -// email_fields['errorReport'] = (workflow.errorReport ?: 'None') -// email_fields['commandLine'] = workflow.commandLine -// email_fields['projectDir'] = workflow.projectDir -// email_fields['summary'] = summary -// email_fields['summary']['Date Started'] = workflow.start -// email_fields['summary']['Date Completed'] = workflow.complete -// email_fields['summary']['Pipeline script file path'] = workflow.scriptFile -// email_fields['summary']['Pipeline script hash ID'] = workflow.scriptId -// if (workflow.repository) email_fields['summary']['Pipeline repository Git URL'] = workflow.repository -// if (workflow.commitId) email_fields['summary']['Pipeline repository Git Commit'] = workflow.commitId -// if (workflow.revision) email_fields['summary']['Pipeline Git branch/tag'] = workflow.revision -// if (workflow.container) email_fields['summary']['Docker image'] = workflow.container -// email_fields['summary']['Nextflow Version'] = workflow.nextflow.version -// email_fields['summary']['Nextflow Build'] = workflow.nextflow.build -// email_fields['summary']['Nextflow Compile Timestamp'] = workflow.nextflow.timestamp -// -// // On success try attach the multiqc report -// def mqc_report = null -// try { -// if (workflow.success) { -// mqc_report = ch_multiqc_report.getVal() -// if (mqc_report.getClass() == ArrayList) { -// log.warn "[nf-core/atacseq] Found multiple reports from process 'multiqc', will use only one" -// mqc_report = mqc_report[0] -// } -// } -// } catch (all) { -// log.warn "[nf-core/atacseq] Could not attach MultiQC report to summary email" -// } -// -// // Check if we are only sending emails on failure -// email_address = params.email -// if (!params.email && params.email_on_fail && !workflow.success) { -// email_address = params.email_on_fail -// } -// -// // Render the TXT template -// def engine = new groovy.text.GStringTemplateEngine() -// def tf = new File("$baseDir/assets/email_template.txt") -// def txt_template = engine.createTemplate(tf).make(email_fields) -// def email_txt = txt_template.toString() -// -// // Render the HTML template -// def hf = new File("$baseDir/assets/email_template.html") -// def html_template = engine.createTemplate(hf).make(email_fields) -// def email_html = html_template.toString() -// -// // Render the sendmail template -// def smail_fields = [ email: email_address, subject: subject, email_txt: email_txt, email_html: email_html, baseDir: "$baseDir", mqcFile: mqc_report, mqcMaxSize: params.max_multiqc_email_size.toBytes() ] -// def sf = new File("$baseDir/assets/sendmail_template.txt") -// def sendmail_template = engine.createTemplate(sf).make(smail_fields) -// def sendmail_html = sendmail_template.toString() -// -// // Send the HTML e-mail -// if (email_address) { -// try { -// if (params.plaintext_email) { throw GroovyException('Send plaintext e-mail, not HTML') } -// // Try to send HTML e-mail using sendmail -// [ 'sendmail', '-t' ].execute() << sendmail_html -// log.info "[nf-core/atacseq] Sent summary e-mail to $email_address (sendmail)" -// } catch (all) { -// // Catch failures and try with plaintext -// [ 'mail', '-s', subject, email_address ].execute() << email_txt -// log.info "[nf-core/atacseq] Sent summary e-mail to $email_address (mail)" -// } -// } -// -// // Write summary e-mail HTML to a file -// def output_d = new File("${params.outdir}/pipeline_info/") -// if (!output_d.exists()) { -// output_d.mkdirs() -// } -// def output_hf = new File(output_d, "pipeline_report.html") -// output_hf.withWriter { w -> w << email_html } -// def output_tf = new File(output_d, "pipeline_report.txt") -// output_tf.withWriter { w -> w << email_txt } -// -// c_reset = params.monochrome_logs ? '' : "\033[0m"; -// c_purple = params.monochrome_logs ? '' : "\033[0;35m"; -// c_green = params.monochrome_logs ? '' : "\033[0;32m"; -// c_red = params.monochrome_logs ? '' : "\033[0;31m"; -// -// if (workflow.stats.ignoredCount > 0 && workflow.success) { -// log.info "${c_purple}Warning, pipeline completed, but with errored process(es) ${c_reset}" -// log.info "${c_red}Number of ignored errored process(es) : ${workflow.stats.ignoredCount} ${c_reset}" -// log.info "${c_green}Number of successfully ran process(es) : ${workflow.stats.succeedCount} ${c_reset}" -// } -// -// if (workflow.success) { -// log.info "${c_purple}[nf-core/atacseq]${c_green} Pipeline completed successfully${c_reset}" -// } else { -// checkHostname() -// log.info "${c_purple}[nf-core/atacseq]${c_red} Pipeline completed with errors${c_reset}" -// } -// -// } +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// +/* -- -- */ +/* -- FASTQ QC -- */ +/* -- -- */ +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// + +/* + * STEP 1 - FastQC + */ +process FastQC { + tag "$name" + label 'process_medium' + publishDir "${params.outdir}/fastqc", mode: 'copy', + saveAs: { filename -> + filename.endsWith(".zip") ? "zips/$filename" : "$filename" + } + + when: + !params.skip_fastqc + + input: + set val(name), file(reads) from ch_raw_reads_fastqc + + output: + file "*.{zip,html}" into ch_fastqc_reports_mqc + + script: + // Added soft-links to original fastqs for consistent naming in MultiQC + if (params.single_end) { + """ + [ ! -f ${name}.fastq.gz ] && ln -s $reads ${name}.fastq.gz + fastqc -q -t $task.cpus ${name}.fastq.gz + """ + } else { + """ + [ ! -f ${name}_1.fastq.gz ] && ln -s ${reads[0]} ${name}_1.fastq.gz + [ ! -f ${name}_2.fastq.gz ] && ln -s ${reads[1]} ${name}_2.fastq.gz + fastqc -q -t $task.cpus ${name}_1.fastq.gz + fastqc -q -t $task.cpus ${name}_2.fastq.gz + """ + } +} + +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// +/* -- -- */ +/* -- ADAPTER TRIMMING -- */ +/* -- -- */ +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// + +/* + * STEP 2 - Trim Galore! + */ +if (params.skip_trimming) { + ch_trimmed_reads = ch_raw_reads_trimgalore + ch_trimgalore_results_mqc = Channel.empty() + ch_trimgalore_fastqc_reports_mqc = Channel.empty() +} else { + process TrimGalore { + tag "$name" + label 'process_high' + publishDir "${params.outdir}/trim_galore", mode: 'copy', + saveAs: { filename -> + if (filename.endsWith(".html")) "fastqc/$filename" + else if (filename.endsWith(".zip")) "fastqc/zips/$filename" + else if (filename.endsWith("trimming_report.txt")) "logs/$filename" + else params.save_trimmed ? filename : null + } + + input: + set val(name), file(reads) from ch_raw_reads_trimgalore + + output: + set val(name), file("*.fq.gz") into ch_trimmed_reads + file "*.txt" into ch_trimgalore_results_mqc + file "*.{zip,html}" into ch_trimgalore_fastqc_reports_mqc + + script: + // Calculate number of --cores for TrimGalore based on value of task.cpus + // See: https://github.com/FelixKrueger/TrimGalore/blob/master/Changelog.md#version-060-release-on-1-mar-2019 + // See: https://github.com/nf-core/atacseq/pull/65 + def cores = 1 + if (task.cpus) { + cores = (task.cpus as int) - 4 + if (params.single_end) cores = (task.cpus as int) - 3 + if (cores < 1) cores = 1 + if (cores > 4) cores = 4 + } + + // Added soft-links to original fastqs for consistent naming in MultiQC + c_r1 = params.clip_r1 > 0 ? "--clip_r1 ${params.clip_r1}" : '' + c_r2 = params.clip_r2 > 0 ? "--clip_r2 ${params.clip_r2}" : '' + tpc_r1 = params.three_prime_clip_r1 > 0 ? "--three_prime_clip_r1 ${params.three_prime_clip_r1}" : '' + tpc_r2 = params.three_prime_clip_r2 > 0 ? "--three_prime_clip_r2 ${params.three_prime_clip_r2}" : '' + nextseq = params.trim_nextseq > 0 ? "--nextseq ${params.trim_nextseq}" : '' + + // Added soft-links to original fastqs for consistent naming in MultiQC + if (params.single_end) { + """ + [ ! -f ${name}.fastq.gz ] && ln -s $reads ${name}.fastq.gz + trim_galore --cores $cores --fastqc --gzip $c_r1 $tpc_r1 $nextseq ${name}.fastq.gz + """ + } else { + """ + [ ! -f ${name}_1.fastq.gz ] && ln -s ${reads[0]} ${name}_1.fastq.gz + [ ! -f ${name}_2.fastq.gz ] && ln -s ${reads[1]} ${name}_2.fastq.gz + trim_galore --cores $cores --paired --fastqc --gzip $c_r1 $c_r2 $tpc_r1 $tpc_r2 $nextseq ${name}_1.fastq.gz ${name}_2.fastq.gz + """ + } + } +} + +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// +/* -- -- */ +/* -- ALIGN -- */ +/* -- -- */ +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// + +/* + * STEP 3.1 - Align read 1 with bwa + */ +process BWAMem { + tag "$name" + label 'process_high' + + input: + set val(name), file(reads) from ch_trimmed_reads + file index from ch_bwa_index.collect() + + output: + set val(name), file("*.bam") into ch_bwa_bam + + script: + prefix = "${name}.Lb" + rg = "\'@RG\\tID:${name}\\tSM:${name.split('_')[0..-2].join('_')}\\tPL:ILLUMINA\\tLB:${name}\\tPU:1\'" + if (params.seq_center) { + rg = "\'@RG\\tID:${name}\\tSM:${name.split('_')[0..-2].join('_')}\\tPL:ILLUMINA\\tLB:${name}\\tPU:1\\tCN:${params.seq_center}\'" + } + """ + bwa mem \\ + -t $task.cpus \\ + -M \\ + -R $rg \\ + ${index}/${bwa_base} \\ + $reads \\ + | samtools view -@ $task.cpus -b -h -F 0x0100 -O BAM -o ${prefix}.bam - + """ +} + +/* + * STEP 3.2 - Convert .bam to coordinate sorted .bam + */ +process SortBAM { + tag "$name" + label 'process_medium' + if (params.save_align_intermeds) { + publishDir path: "${params.outdir}/bwa/library", mode: 'copy', + saveAs: { filename -> + if (filename.endsWith(".flagstat")) "samtools_stats/$filename" + else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" + else if (filename.endsWith(".stats")) "samtools_stats/$filename" + else filename + } + } + + input: + set val(name), file(bam) from ch_bwa_bam + + output: + set val(name), file("*.sorted.{bam,bam.bai}") into ch_sort_bam_merge + file "*.{flagstat,idxstats,stats}" into ch_sort_bam_flagstat_mqc + + script: + prefix = "${name}.Lb" + """ + samtools sort -@ $task.cpus -o ${prefix}.sorted.bam -T $name $bam + samtools index ${prefix}.sorted.bam + samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat + samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats + samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats + """ +} + +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// +/* -- -- */ +/* -- MERGE LIBRARY BAM -- */ +/* -- -- */ +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// + +/* + * STEP 4.1 Merge BAM files for all libraries from same replicate + */ +ch_sort_bam_merge + .map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] } + .groupTuple(by: [0]) + .map { it -> [ it[0], it[1].flatten() ] } + .set { ch_sort_bam_merge } + +process MergedLibBAM { + tag "$name" + label 'process_medium' + publishDir "${params.outdir}/bwa/mergedLibrary", mode: 'copy', + saveAs: { filename -> + if (filename.endsWith(".flagstat")) "samtools_stats/$filename" + else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" + else if (filename.endsWith(".stats")) "samtools_stats/$filename" + else if (filename.endsWith(".metrics.txt")) "picard_metrics/$filename" + else params.save_align_intermeds ? filename : null + } + + input: + set val(name), file(bams) from ch_sort_bam_merge + + output: + set val(name), file("*${prefix}.sorted.{bam,bam.bai}") into ch_mlib_bam_filter, + ch_mlib_bam_preseq, + ch_mlib_bam_ataqv + file "*.{flagstat,idxstats,stats}" into ch_mlib_bam_stats_mqc + file "*.txt" into ch_mlib_bam_metrics_mqc + + script: + prefix = "${name}.mLb.mkD" + bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() + def avail_mem = 3 + if (!task.memory) { + log.info "[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." + } else { + avail_mem = task.memory.toGiga() + } + if (bam_files.size() > 1) { + """ + picard -Xmx${avail_mem}g MergeSamFiles \\ + ${'INPUT='+bam_files.join(' INPUT=')} \\ + OUTPUT=${name}.sorted.bam \\ + SORT_ORDER=coordinate \\ + VALIDATION_STRINGENCY=LENIENT \\ + TMP_DIR=tmp + samtools index ${name}.sorted.bam + + picard -Xmx${avail_mem}g MarkDuplicates \\ + INPUT=${name}.sorted.bam \\ + OUTPUT=${prefix}.sorted.bam \\ + ASSUME_SORTED=true \\ + REMOVE_DUPLICATES=false \\ + METRICS_FILE=${prefix}.MarkDuplicates.metrics.txt \\ + VALIDATION_STRINGENCY=LENIENT \\ + TMP_DIR=tmp + + samtools index ${prefix}.sorted.bam + samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats + samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat + samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats + """ + } else { + """ + picard -Xmx${avail_mem}g MarkDuplicates \\ + INPUT=${bam_files[0]} \\ + OUTPUT=${prefix}.sorted.bam \\ + ASSUME_SORTED=true \\ + REMOVE_DUPLICATES=false \\ + METRICS_FILE=${prefix}.MarkDuplicates.metrics.txt \\ + VALIDATION_STRINGENCY=LENIENT \\ + TMP_DIR=tmp + + samtools index ${prefix}.sorted.bam + samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats + samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat + samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats + """ + } +} + +/* + * STEP 4.2 Filter BAM file at merged library-level + */ +process MergedLibBAMFilter { + tag "$name" + label 'process_medium' + publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', + saveAs: { filename -> + if (params.single_end || params.save_align_intermeds) { + if (filename.endsWith(".flagstat")) "samtools_stats/$filename" + else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" + else if (filename.endsWith(".stats")) "samtools_stats/$filename" + else if (filename.endsWith(".sorted.bam")) filename + else if (filename.endsWith(".sorted.bam.bai")) filename + else null + } + } + + input: + set val(name), file(bam) from ch_mlib_bam_filter + file bed from ch_genome_filter_regions.collect() + file bamtools_filter_config from ch_bamtools_filter_config + + output: + set val(name), file("*.{bam,bam.bai}") into ch_mlib_filter_bam + set val(name), file("*.flagstat") into ch_mlib_filter_bam_flagstat + file "*.{idxstats,stats}" into ch_mlib_filter_bam_stats_mqc + + script: + prefix = params.single_end ? "${name}.mLb.clN" : "${name}.mLb.flT" + filter_params = params.single_end ? "-F 0x004" : "-F 0x004 -F 0x0008 -f 0x001" + dup_params = params.keep_dups ? "" : "-F 0x0400" + multimap_params = params.keep_multi_map ? "" : "-q 1" + blacklist_params = params.blacklist ? "-L $bed" : "" + name_sort_bam = params.single_end ? "" : "samtools sort -n -@ $task.cpus -o ${prefix}.bam -T $prefix ${prefix}.sorted.bam" + """ + samtools view \\ + $filter_params \\ + $dup_params \\ + $multimap_params \\ + $blacklist_params \\ + -b ${bam[0]} \\ + | bamtools filter \\ + -out ${prefix}.sorted.bam \\ + -script $bamtools_filter_config + + samtools index ${prefix}.sorted.bam + samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat + samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats + samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats + + $name_sort_bam + """ +} + +/* + * STEP 4.3 Remove orphan reads from paired-end BAM file + */ +if (params.single_end) { + ch_mlib_filter_bam + .into { ch_mlib_rm_orphan_bam_metrics; + ch_mlib_rm_orphan_bam_bigwig; + ch_mlib_rm_orphan_bam_macs; + ch_mlib_rm_orphan_bam_plotfingerprint; + ch_mlib_rm_orphan_bam_mrep; + ch_mlib_name_bam_mlib_counts; + ch_mlib_name_bam_mrep_counts } + + ch_mlib_filter_bam_flagstat + .into { ch_mlib_rm_orphan_flagstat_bigwig; + ch_mlib_rm_orphan_flagstat_macs; + ch_mlib_rm_orphan_flagstat_mqc } + + ch_mlib_filter_bam_stats_mqc + .set { ch_mlib_rm_orphan_stats_mqc } +} else { + process MergedLibBAMRemoveOrphan { + tag "$name" + label 'process_medium' + publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', + saveAs: { filename -> + if (filename.endsWith(".flagstat")) "samtools_stats/$filename" + else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" + else if (filename.endsWith(".stats")) "samtools_stats/$filename" + else if (filename.endsWith(".sorted.bam")) filename + else if (filename.endsWith(".sorted.bam.bai")) filename + else null + } + + input: + set val(name), file(bam) from ch_mlib_filter_bam + + output: + set val(name), file("*.sorted.{bam,bam.bai}") into ch_mlib_rm_orphan_bam_metrics, + ch_mlib_rm_orphan_bam_bigwig, + ch_mlib_rm_orphan_bam_macs, + ch_mlib_rm_orphan_bam_plotfingerprint, + ch_mlib_rm_orphan_bam_mrep + set val(name), file("${prefix}.bam") into ch_mlib_name_bam_mlib_counts, + ch_mlib_name_bam_mrep_counts + set val(name), file("*.flagstat") into ch_mlib_rm_orphan_flagstat_bigwig, + ch_mlib_rm_orphan_flagstat_macs, + ch_mlib_rm_orphan_flagstat_mqc + file "*.{idxstats,stats}" into ch_mlib_rm_orphan_stats_mqc + + script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ + prefix = "${name}.mLb.clN" + """ + bampe_rm_orphan.py ${bam[0]} ${prefix}.bam --only_fr_pairs + + samtools sort -@ $task.cpus -o ${prefix}.sorted.bam -T $prefix ${prefix}.bam + samtools index ${prefix}.sorted.bam + samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat + samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats + samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats + """ + } +} + +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// +/* -- -- */ +/* -- MERGE LIBRARY BAM POST-ANALYSIS -- */ +/* -- -- */ +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// + +/* + * STEP 5.1 preseq analysis after merging libraries and before filtering + */ +process MergedLibPreseq { + tag "$name" + label 'process_low' + publishDir "${params.outdir}/bwa/mergedLibrary/preseq", mode: 'copy' + + when: + !params.skip_preseq + + input: + set val(name), file(bam) from ch_mlib_bam_preseq + + output: + file "*.ccurve.txt" into ch_mlib_preseq_mqc + + script: + prefix = "${name}.mLb.mkD" + """ + preseq lc_extrap -v -output ${prefix}.ccurve.txt -bam ${bam[0]} + """ +} + +/* + * STEP 5.2 Picard CollectMultipleMetrics after merging libraries and filtering + */ +process MergedLibMetrics { + tag "$name" + label 'process_medium' + publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', + saveAs: { filename -> + if (filename.endsWith("_metrics")) "picard_metrics/$filename" + else if (filename.endsWith(".pdf")) "picard_metrics/pdf/$filename" + else null + } + + when: + !params.skip_picard_metrics + + input: + set val(name), file(bam) from ch_mlib_rm_orphan_bam_metrics + file fasta from ch_fasta + + output: + file "*_metrics" into ch_mlib_collectmetrics_mqc + file "*.pdf" into ch_mlib_collectmetrics_pdf + + script: + prefix = "${name}.mLb.clN" + def avail_mem = 3 + if (!task.memory) { + log.info "[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." + } else { + avail_mem = task.memory.toGiga() + } + """ + picard -Xmx${avail_mem}g CollectMultipleMetrics \\ + INPUT=${bam[0]} \\ + OUTPUT=${prefix}.CollectMultipleMetrics \\ + REFERENCE_SEQUENCE=$fasta \\ + VALIDATION_STRINGENCY=LENIENT \\ + TMP_DIR=tmp + """ +} + +/* + * STEP 5.3 Read depth normalised bigWig + */ +process MergedLibBigWig { + tag "$name" + label 'process_medium' + publishDir "${params.outdir}/bwa/mergedLibrary/bigwig", mode: 'copy', + saveAs: { filename -> + if (filename.endsWith("scale_factor.txt")) "scale/$filename" + else if (filename.endsWith(".bigWig")) "$filename" + else null + } + + input: + set val(name), file(bam), file(flagstat) from ch_mlib_rm_orphan_bam_bigwig.join(ch_mlib_rm_orphan_flagstat_bigwig, by: [0]) + file sizes from ch_genome_sizes_mlib_bigwig.collect() + + output: + set val(name), file("*.bigWig") into ch_mlib_bigwig_plotprofile + file "*scale_factor.txt" into ch_mlib_bigwig_scale + file "*igv.txt" into ch_mlib_bigwig_igv + + script: + prefix = "${name}.mLb.clN" + pe_fragment = params.single_end ? "" : "-pc" + extend = (params.single_end && params.fragment_size > 0) ? "-fs ${params.fragment_size}" : '' + """ + SCALE_FACTOR=\$(grep 'mapped (' $flagstat | awk '{print 1000000/\$1}') + echo \$SCALE_FACTOR > ${prefix}.scale_factor.txt + genomeCoverageBed -ibam ${bam[0]} -bg -scale \$SCALE_FACTOR $pe_fragment $extend | sort -T '.' -k1,1 -k2,2n > ${prefix}.bedGraph + + bedGraphToBigWig ${prefix}.bedGraph $sizes ${prefix}.bigWig + + find * -type f -name "*.bigWig" -exec echo -e "bwa/mergedLibrary/bigwig/"{}"\\t0,0,178" \\; > ${prefix}.bigWig.igv.txt + """ +} + +/* + * STEP 5.4 generate gene body coverage plot with deepTools + */ +process MergedLibPlotProfile { + tag "$name" + label 'process_high' + publishDir "${params.outdir}/bwa/mergedLibrary/deepTools/plotProfile", mode: 'copy' + + when: + !params.skip_plot_profile + + input: + set val(name), file(bigwig) from ch_mlib_bigwig_plotprofile + file bed from ch_gene_bed + + output: + file '*.{gz,pdf}' into ch_mlib_plotprofile_results + file '*.plotProfile.tab' into ch_mlib_plotprofile_mqc + + script: + prefix = "${name}.mLb.clN" + """ + computeMatrix scale-regions \\ + --regionsFileName $bed \\ + --scoreFileName $bigwig \\ + --outFileName ${prefix}.computeMatrix.mat.gz \\ + --outFileNameMatrix ${prefix}.computeMatrix.vals.mat.gz \\ + --regionBodyLength 1000 \\ + --beforeRegionStartLength 3000 \\ + --afterRegionStartLength 3000 \\ + --skipZeros \\ + --smartLabels \\ + --numberOfProcessors $task.cpus + + plotProfile --matrixFile ${prefix}.computeMatrix.mat.gz \\ + --outFileName ${prefix}.plotProfile.pdf \\ + --outFileNameData ${prefix}.plotProfile.tab + """ +} + +/* + * STEP 5.5 deepTools plotFingerprint + */ +process MergedLibPlotFingerprint { + tag "$name" + label 'process_high' + publishDir "${params.outdir}/bwa/mergedLibrary/deepTools/plotFingerprint", mode: 'copy' + + when: + !params.skip_plot_fingerprint + + input: + set val(name), file(bam) from ch_mlib_rm_orphan_bam_plotfingerprint + + output: + file '*.{txt,pdf}' into ch_mlib_plotfingerprint_results + file '*.raw.txt' into ch_mlib_plotfingerprint_mqc + + script: + prefix = "${name}.mLb.clN" + extend = (params.single_end && params.fragment_size > 0) ? "--extendReads ${params.fragment_size}" : '' + """ + plotFingerprint \\ + --bamfiles ${bam[0]} \\ + --plotFile ${prefix}.plotFingerprint.pdf \\ + $extend \\ + --labels $prefix \\ + --outRawCounts ${prefix}.plotFingerprint.raw.txt \\ + --outQualityMetrics ${prefix}.plotFingerprint.qcmetrics.txt \\ + --skipZeros \\ + --numberOfProcessors $task.cpus \\ + --numberOfSamples $params.fingerprint_bins + """ +} + +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// +/* -- -- */ +/* -- MERGE LIBRARY PEAK ANALYSIS -- */ +/* -- -- */ +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// + +/* + * STEP 6.1 Call peaks with MACS2 and calculate FRiP score + */ +process MergedLibMACSCallPeak { + tag "$name" + label 'process_medium' + publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}", mode: 'copy', + saveAs: { filename -> + if (filename.endsWith(".tsv")) "qc/$filename" + else if (filename.endsWith(".igv.txt")) null + else filename + } + + when: + params.macs_gsize + + input: + set val(name), file(bam), file(flagstat) from ch_mlib_rm_orphan_bam_macs.join(ch_mlib_rm_orphan_flagstat_macs, by: [0]) + file mlib_peak_count_header from ch_mlib_peak_count_header + file mlib_frip_score_header from ch_mlib_frip_score_header + + output: + set val(name), file("*.{bed,xls,gappedPeak,bdg}") into ch_mlib_macs_output + set val(name), file("*$PEAK_TYPE") into ch_mlib_macs_homer, + ch_mlib_macs_qc, + ch_mlib_macs_consensus, + ch_mlib_macs_ataqv + file "*igv.txt" into ch_mlib_macs_igv + file "*_mqc.tsv" into ch_mlib_macs_mqc + + script: + prefix = "${name}.mLb.clN" + broad = params.narrow_peak ? '' : "--broad --broad-cutoff ${params.broad_cutoff}" + format = params.single_end ? "BAM" : "BAMPE" + pileup = params.save_macs_pileup ? "-B --SPMR" : "" + """ + macs2 callpeak \\ + -t ${bam[0]} \\ + $broad \\ + -f $format \\ + -g $params.macs_gsize \\ + -n $prefix \\ + $pileup \\ + --keep-dup all \\ + --nomodel + + cat ${prefix}_peaks.${PEAK_TYPE} | wc -l | awk -v OFS='\t' '{ print "${name}", \$1 }' | cat $mlib_peak_count_header - > ${prefix}_peaks.count_mqc.tsv + + READS_IN_PEAKS=\$(intersectBed -a ${bam[0]} -b ${prefix}_peaks.${PEAK_TYPE} -bed -c -f 0.20 | awk -F '\t' '{sum += \$NF} END {print sum}') + grep 'mapped (' $flagstat | awk -v a="\$READS_IN_PEAKS" -v OFS='\t' '{print "${name}", a/\$1}' | cat $mlib_frip_score_header - > ${prefix}_peaks.FRiP_mqc.tsv + + find * -type f -name "*.${PEAK_TYPE}" -exec echo -e "bwa/mergedLibrary/macs/${PEAK_TYPE}/"{}"\\t0,0,178" \\; > ${prefix}_peaks.igv.txt + """ +} + +/* + * STEP 6.2 Annotate peaks with HOMER + */ +process MergedLibAnnotatePeaks { + tag "$name" + label "process_medium" + publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}", mode: 'copy' + + when: + params.macs_gsize + + input: + set val(name), file(peak) from ch_mlib_macs_homer + file fasta from ch_fasta + file gtf from ch_gtf + + output: + file "*.txt" into ch_mlib_macs_annotate + + script: + prefix = "${name}.mLb.clN" + """ + annotatePeaks.pl \\ + $peak \\ + $fasta \\ + -gid \\ + -gtf $gtf \\ + -cpu $task.cpus \\ + > ${prefix}_peaks.annotatePeaks.txt + """ +} + +/* + * STEP 6.3 Aggregated QC plots for peaks, FRiP and peak-to-gene annotation + */ +process MergedLibPeakQC { + label "process_medium" + publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/qc", mode: 'copy' + + when: + params.macs_gsize + + input: + file peaks from ch_mlib_macs_qc.collect{ it[1] } + file annos from ch_mlib_macs_annotate.collect() + file mlib_peak_annotation_header from ch_mlib_peak_annotation_header + + output: + file "*.{txt,pdf}" into ch_mlib_peak_qc + file "*.tsv" into ch_mlib_peak_qc_mqc + + script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ + suffix = 'mLb.clN' + """ + plot_macs_qc.r \\ + -i ${peaks.join(',')} \\ + -s ${peaks.join(',').replaceAll(".${suffix}_peaks.${PEAK_TYPE}","")} \\ + -o ./ \\ + -p macs_peak.${suffix} + + plot_homer_annotatepeaks.r \\ + -i ${annos.join(',')} \\ + -s ${annos.join(',').replaceAll(".${suffix}_peaks.annotatePeaks.txt","")} \\ + -o ./ \\ + -p macs_annotatePeaks.${suffix} + + cat $mlib_peak_annotation_header macs_annotatePeaks.${suffix}.summary.txt > macs_annotatePeaks.${suffix}.summary_mqc.tsv + """ +} + +/* + * STEP 6.4 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection + */ +process MergedLibConsensusPeakSet { + label 'process_long' + publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: 'copy', + saveAs: { filename -> + if (filename.endsWith(".igv.txt")) null + else filename + } + + when: + params.macs_gsize && (replicatesExist || multipleGroups) && !params.skip_consensus_peaks + + input: + file peaks from ch_mlib_macs_consensus.collect{ it[1] } + + output: + file "*.bed" into ch_mlib_macs_consensus_bed + file "*.saf" into ch_mlib_macs_consensus_saf + file "*.boolean.txt" into ch_mlib_macs_consensus_bool + file "*.intersect.{txt,plot.pdf}" into ch_mlib_macs_consensus_intersect + file "*igv.txt" into ch_mlib_macs_consensus_igv + + script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ + suffix = 'mLb.clN' + prefix = "consensus_peaks.${suffix}" + mergecols = params.narrow_peak ? (2..10).join(',') : (2..9).join(',') + collapsecols = params.narrow_peak ? (["collapse"]*9).join(',') : (["collapse"]*8).join(',') + expandparam = params.narrow_peak ? "--is_narrow_peak" : "" + """ + sort -T '.' -k1,1 -k2,2n ${peaks.collect{it.toString()}.sort().join(' ')} \\ + | mergeBed -c $mergecols -o $collapsecols > ${prefix}.txt + + macs2_merged_expand.py \\ + ${prefix}.txt \\ + ${peaks.collect{it.toString()}.sort().join(',').replaceAll("_peaks.${PEAK_TYPE}","")} \\ + ${prefix}.boolean.txt \\ + --min_replicates $params.min_reps_consensus \\ + $expandparam + + awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$1, \$2, \$3, \$4, "0", "+" }' ${prefix}.boolean.txt > ${prefix}.bed + + echo -e "GeneID\tChr\tStart\tEnd\tStrand" > ${prefix}.saf + awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$4, \$1, \$2, \$3, "+" }' ${prefix}.boolean.txt >> ${prefix}.saf + + sed -i 's/.${suffix}//g' ${prefix}.boolean.intersect.txt + plot_peak_intersect.r -i ${prefix}.boolean.intersect.txt -o ${prefix}.boolean.intersect.plot.pdf + + find * -type f -name "${prefix}.bed" -exec echo -e "bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus/"{}"\\t0,0,0" \\; > ${prefix}.bed.igv.txt + """ +} + +/* + * STEP 6.5 Annotate consensus peaks with HOMER, and add annotation to boolean output file + */ +process MergedLibConsensusPeakSetAnnotate { + label "process_medium" + publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: 'copy' + + when: + params.macs_gsize && (replicatesExist || multipleGroups) && !params.skip_consensus_peaks + + input: + file bed from ch_mlib_macs_consensus_bed + file bool from ch_mlib_macs_consensus_bool + file fasta from ch_fasta + file gtf from ch_gtf + + output: + file "*.annotatePeaks.txt" into ch_mlib_macs_consensus_annotate + + script: + prefix = "consensus_peaks.mLb.clN" + """ + annotatePeaks.pl \\ + $bed \\ + $fasta \\ + -gid \\ + -gtf $gtf \\ + -cpu $task.cpus \\ + > ${prefix}.annotatePeaks.txt + + cut -f2- ${prefix}.annotatePeaks.txt | awk 'NR==1; NR > 1 {print \$0 | "sort -T '.' -k1,1 -k2,2n"}' | cut -f6- > tmp.txt + paste $bool tmp.txt > ${prefix}.boolean.annotatePeaks.txt + """ +} + +/* + * STEP 6.6 Count reads in consensus peaks with featureCounts and perform differential analysis with DESeq2 + */ +process MergedLibConsensusPeakSetDESeq { + label 'process_medium' + publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus/deseq2", mode: 'copy', + saveAs: { filename -> + if (filename.endsWith(".igv.txt")) null + else filename + } + + when: + params.macs_gsize && replicatesExist && multipleGroups && !params.skip_consensus_peaks + + input: + file bams from ch_mlib_name_bam_mlib_counts.collect{ it[1] } + file saf from ch_mlib_macs_consensus_saf.collect() + file mlib_deseq2_pca_header from ch_mlib_deseq2_pca_header + file mlib_deseq2_clustering_header from ch_mlib_deseq2_clustering_header + + output: + file "*featureCounts.txt" into ch_mlib_macs_consensus_counts + file "*featureCounts.txt.summary" into ch_mlib_macs_consensus_counts_mqc + file "*.{RData,results.txt,pdf,log}" into ch_mlib_macs_consensus_deseq_results + file "sizeFactors" into ch_mlib_macs_consensus_deseq_factors + file "*vs*/*.{pdf,txt}" into ch_mlib_macs_consensus_deseq_comp_results + file "*vs*/*.bed" into ch_mlib_macs_consensus_deseq_comp_bed + file "*igv.txt" into ch_mlib_macs_consensus_deseq_comp_igv + file "*.tsv" into ch_mlib_macs_consensus_deseq_mqc + + script: + prefix = "consensus_peaks.mLb.clN" + bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() + bam_ext = params.single_end ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" + pe_params = params.single_end ? '' : "-p --donotsort" + """ + featureCounts \\ + -F SAF \\ + -O \\ + --fracOverlap 0.2 \\ + -T $task.cpus \\ + $pe_params \\ + -a $saf \\ + -o ${prefix}.featureCounts.txt \\ + ${bam_files.join(' ')} + + featurecounts_deseq2.r -i ${prefix}.featureCounts.txt -b '$bam_ext' -o ./ -p $prefix -s .mLb + + cat $mlib_deseq2_pca_header ${prefix}.pca.vals.txt > ${prefix}.pca.vals_mqc.tsv + cat $mlib_deseq2_clustering_header ${prefix}.sample.dists.txt > ${prefix}.sample.dists_mqc.tsv + + find * -type f -name "*.FDR0.05.results.bed" -exec echo -e "bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus/deseq2/"{}"\\t255,0,0" \\; > ${prefix}.igv.txt + """ +} + +/* + * STEP 6.7 Run ataqv on BAM file and corresponding peaks + */ +process MergedLibAtaqv { + tag "$name" + label 'process_medium' + publishDir "${params.outdir}/bwa/mergedLibrary/ataqv/${PEAK_TYPE}", mode: 'copy' + + when: + !params.skip_ataqv + + input: + set val(name), file(bam), file(peak) from ch_mlib_bam_ataqv.join(ch_mlib_macs_ataqv, by: [0]) + file autosomes from ch_genome_autosomes.collect() + file tss_bed from ch_tss_bed + + output: + file "*.json" into ch_mlib_ataqv + + script: + peak_param = params.macs_gsize ? "--peak-file ${peak}" : "" + mito_param = params.mito_name ? "--mitochondrial-reference-name ${params.mito_name}" : "" + """ + ataqv \\ + --threads $task.cpus \\ + $peak_param \\ + --tss-file $tss_bed \\ + --metrics-file ${name}.ataqv.json \\ + --name $name \\ + --ignore-read-groups \\ + --autosomal-reference-file $autosomes \\ + $mito_param \\ + NA \\ + ${bam[0]} + """ +} + +/* + * STEP 6.8 run ataqv mkarv on all JSON files to render web app + */ +process MergedLibAtaqvMkarv { + label 'process_medium' + publishDir "${params.outdir}/bwa/mergedLibrary/ataqv/${PEAK_TYPE}", mode: 'copy' + + when: + !params.skip_ataqv + + input: + file json from ch_mlib_ataqv.collect() + + output: + file "html" into ch_mlib_ataqv_mkarv + + script: + """ + mkarv \\ + --concurrency $task.cpus \\ + --force \\ + ./html/ \\ + ${json.join(' ')} + """ +} + +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// +/* -- -- */ +/* -- MERGE REPLICATE BAM -- */ +/* -- -- */ +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// + +/* + * STEP 7 Merge library BAM files across all replicates + */ +ch_mlib_rm_orphan_bam_mrep + .map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] } + .groupTuple(by: [0]) + .map { it -> [ it[0], it[1].flatten() ] } + .set { ch_mlib_rm_orphan_bam_mrep } + +process MergedRepBAM { + tag "$name" + label 'process_medium' + publishDir "${params.outdir}/bwa/mergedReplicate", mode: 'copy', + saveAs: { filename -> + if (filename.endsWith(".flagstat")) "samtools_stats/$filename" + else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" + else if (filename.endsWith(".stats")) "samtools_stats/$filename" + else if (filename.endsWith(".metrics.txt")) "picard_metrics/$filename" + else filename + } + + input: + set val(name), file(bams) from ch_mlib_rm_orphan_bam_mrep + + output: + set val(name), file("*${prefix}.sorted.{bam,bam.bai}") into ch_mrep_bam_bigwig, + ch_mrep_bam_macs + set val(name), file("*.flagstat") into ch_mrep_bam_flagstat_bigwig, + ch_mrep_bam_flagstat_macs, + ch_mrep_bam_flagstat_mqc + file "*.{idxstats,stats}" into ch_mrep_bam_stats_mqc + file "*.txt" into ch_mrep_bam_metrics_mqc + + when: + !params.skip_merge_replicates && replicatesExist + + script: + prefix = "${name}.mRp.clN" + bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() + def avail_mem = 3 + if (!task.memory) { + log.info "[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." + } else { + avail_mem = task.memory.toGiga() + } + if (bam_files.size() > 1) { + """ + picard -Xmx${avail_mem}g MergeSamFiles \\ + ${'INPUT='+bam_files.join(' INPUT=')} \\ + OUTPUT=${name}.sorted.bam \\ + SORT_ORDER=coordinate \\ + VALIDATION_STRINGENCY=LENIENT \\ + TMP_DIR=tmp + samtools index ${name}.sorted.bam + + picard -Xmx${avail_mem}g MarkDuplicates \\ + INPUT=${name}.sorted.bam \\ + OUTPUT=${prefix}.sorted.bam \\ + ASSUME_SORTED=true \\ + REMOVE_DUPLICATES=true \\ + METRICS_FILE=${prefix}.MarkDuplicates.metrics.txt \\ + VALIDATION_STRINGENCY=LENIENT \\ + TMP_DIR=tmp + + samtools index ${prefix}.sorted.bam + samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat + samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats + samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats + """ + } else { + """ + ln -s ${bams[0]} ${prefix}.sorted.bam + ln -s ${bams[1]} ${prefix}.sorted.bam.bai + touch ${prefix}.MarkDuplicates.metrics.txt + samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat + samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats + samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats + """ + } +} + +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// +/* -- -- */ +/* -- MERGE REPLICATE BAM POST-ANALYSIS -- */ +/* -- -- */ +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// + +/* + * STEP 8.1 Read depth normalised bigWig + */ +process MergedRepBigWig { + tag "$name" + label 'process_medium' + publishDir "${params.outdir}/bwa/mergedReplicate/bigwig", mode: 'copy', + saveAs: { filename -> + if (filename.endsWith("scale_factor.txt")) "scale/$filename" + else if (filename.endsWith(".bigWig")) "$filename" + else null + } + + when: + !params.skip_merge_replicates && replicatesExist + + input: + set val(name), file(bam), file(flagstat) from ch_mrep_bam_bigwig.join(ch_mrep_bam_flagstat_bigwig, by: [0]) + file sizes from ch_genome_sizes_mrep_bigwig.collect() + + output: + set val(name), file("*.bigWig") into ch_mrep_bigwig + file "*scale_factor.txt" into ch_mrep_bigwig_scale + file "*igv.txt" into ch_mrep_bigwig_igv + + script: + prefix = "${name}.mRp.clN" + pe_fragment = params.single_end ? "" : "-pc" + extend = (params.single_end && params.fragment_size > 0) ? "-fs ${params.fragment_size}" : '' + """ + SCALE_FACTOR=\$(grep 'mapped (' $flagstat | awk '{print 1000000/\$1}') + echo \$SCALE_FACTOR > ${prefix}.scale_factor.txt + genomeCoverageBed -ibam ${bam[0]} -bg -scale \$SCALE_FACTOR $pe_fragment $extend | sort -T '.' -k1,1 -k2,2n > ${prefix}.bedGraph + + bedGraphToBigWig ${prefix}.bedGraph $sizes ${prefix}.bigWig + + find * -type f -name "*.bigWig" -exec echo -e "bwa/mergedReplicate/bigwig/"{}"\\t0,0,178" \\; > ${prefix}.bigWig.igv.txt + """ +} + +/* + * STEP 8.2 Call peaks with MACS2 and calculate FRiP score + */ +process MergedRepMACSCallPeak { + tag "$name" + label 'process_medium' + publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}", mode: 'copy', + saveAs: { filename -> + if (filename.endsWith(".tsv")) "qc/$filename" + else if (filename.endsWith(".igv.txt")) null + else filename + } + + when: + !params.skip_merge_replicates && replicatesExist && params.macs_gsize + + input: + set val(name), file(bam), file(flagstat) from ch_mrep_bam_macs.join(ch_mrep_bam_flagstat_macs, by: [0]) + file mrep_peak_count_header from ch_mrep_peak_count_header + file mrep_frip_score_header from ch_mrep_frip_score_header + + output: + set val(name), file("*.{bed,xls,gappedPeak,bdg}") into ch_mrep_macs_output + set val(name), file("*$PEAK_TYPE") into ch_mrep_macs_homer, + ch_mrep_macs_qc, + ch_mrep_macs_consensus + file "*igv.txt" into ch_mrep_macs_igv + file "*_mqc.tsv" into ch_mrep_macs_mqc + + script: + prefix = "${name}.mRp.clN" + broad = params.narrow_peak ? '' : "--broad --broad-cutoff ${params.broad_cutoff}" + format = params.single_end ? "BAM" : "BAMPE" + pileup = params.save_macs_pileup ? "-B --SPMR" : "" + """ + macs2 callpeak \\ + -t ${bam[0]} \\ + $broad \\ + -f $format \\ + -g $params.macs_gsize \\ + -n $prefix \\ + $pileup \\ + --keep-dup all \\ + --nomodel + + cat ${prefix}_peaks.${PEAK_TYPE} | wc -l | awk -v OFS='\t' '{ print "${name}", \$1 }' | cat $mrep_peak_count_header - > ${prefix}_peaks.count_mqc.tsv + + READS_IN_PEAKS=\$(intersectBed -a ${bam[0]} -b ${prefix}_peaks.${PEAK_TYPE} -bed -c -f 0.20 | awk -F '\t' '{sum += \$NF} END {print sum}') + grep 'mapped (' $flagstat | awk -v a="\$READS_IN_PEAKS" -v OFS='\t' '{print "${name}", a/\$1}' | cat $mrep_frip_score_header - > ${prefix}_peaks.FRiP_mqc.tsv + + find * -type f -name "*.${PEAK_TYPE}" -exec echo -e "bwa/mergedReplicate/macs/${PEAK_TYPE}/"{}"\\t0,0,178" \\; > ${prefix}_peaks.igv.txt + """ +} + +/* + * STEP 8.3 Annotate peaks with HOMER + */ +process MergedRepAnnotatePeaks { + tag "$name" + label "process_medium" + publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}", mode: 'copy' + + when: + !params.skip_merge_replicates && replicatesExist && params.macs_gsize + + input: + set val(name), file(peak) from ch_mrep_macs_homer + file fasta from ch_fasta + file gtf from ch_gtf + + output: + file "*.txt" into ch_mrep_macs_annotate + + script: + prefix = "${name}.mRp.clN" + """ + annotatePeaks.pl \\ + $peak \\ + $fasta \\ + -gid \\ + -gtf $gtf \\ + -cpu $task.cpus \\ + > ${prefix}_peaks.annotatePeaks.txt + """ +} + +/* + * STEP 8.4 Aggregated QC plots for peaks, FRiP and peak-to-gene annotation + */ +process MergedRepPeakQC { + label "process_medium" + publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/qc", mode: 'copy' + + when: + !params.skip_merge_replicates && replicatesExist && params.macs_gsize + + input: + file peaks from ch_mrep_macs_qc.collect{ it[1] } + file annos from ch_mrep_macs_annotate.collect() + file mrep_peak_annotation_header from ch_mrep_peak_annotation_header + + output: + file "*.{txt,pdf}" into ch_mrep_peak_qc + file "*.tsv" into ch_mrep_peak_qc_mqc + + script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ + suffix = 'mRp.clN' + """ + plot_macs_qc.r \\ + -i ${peaks.join(',')} \\ + -s ${peaks.join(',').replaceAll(".${suffix}_peaks.${PEAK_TYPE}","")} \\ + -o ./ \\ + -p macs_peak.${suffix} + + plot_homer_annotatepeaks.r \\ + -i ${annos.join(',')} \\ + -s ${annos.join(',').replaceAll(".${suffix}_peaks.annotatePeaks.txt","")} \\ + -o ./ \\ + -p macs_annotatePeaks.${suffix} + + cat $mrep_peak_annotation_header macs_annotatePeaks.${suffix}.summary.txt > macs_annotatePeaks.${suffix}.summary_mqc.tsv + """ +} + +/* + * STEP 8.5 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection + */ +process MergedRepConsensusPeakSet { + label 'process_long' + publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: 'copy', + saveAs: { filename -> + if (filename.endsWith(".igv.txt")) null + else filename + } + + when: + !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks + + input: + file peaks from ch_mrep_macs_consensus.collect{ it[1] } + + output: + file "*.bed" into ch_mrep_macs_consensus_bed + file "*.saf" into ch_mrep_macs_consensus_saf + file "*.boolean.txt" into ch_mrep_macs_consensus_bool + file "*.intersect.{txt,plot.pdf}" into ch_mrep_macs_consensus_intersect + file "*igv.txt" into ch_mrep_macs_consensus_igv + + script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ + suffix = 'mRp.clN' + prefix = "consensus_peaks.${suffix}" + mergecols = params.narrow_peak ? (2..10).join(',') : (2..9).join(',') + collapsecols = params.narrow_peak ? (["collapse"]*9).join(',') : (["collapse"]*8).join(',') + expandparam = params.narrow_peak ? "--is_narrow_peak" : "" + """ + sort -T '.' -k1,1 -k2,2n ${peaks.collect{it.toString()}.sort().join(' ')} \\ + | mergeBed -c $mergecols -o $collapsecols > ${prefix}.txt + + macs2_merged_expand.py \\ + ${prefix}.txt \\ + ${peaks.collect{it.toString()}.sort().join(',').replaceAll("_peaks.${PEAK_TYPE}","")} \\ + ${prefix}.boolean.txt \\ + $expandparam + + awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$1, \$2, \$3, \$4, "0", "+" }' ${prefix}.boolean.txt > ${prefix}.bed + + echo -e "GeneID\tChr\tStart\tEnd\tStrand" > ${prefix}.saf + awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$4, \$1, \$2, \$3, "+" }' ${prefix}.boolean.txt >> ${prefix}.saf + + sed -i 's/.${suffix}//g' ${prefix}.boolean.intersect.txt + plot_peak_intersect.r -i ${prefix}.boolean.intersect.txt -o ${prefix}.boolean.intersect.plot.pdf + + find * -type f -name "${prefix}.bed" -exec echo -e "bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus/"{}"\\t0,0,0" \\; > ${prefix}.bed.igv.txt + """ +} + +/* + * STEP 8.6 Annotate consensus peaks with HOMER, and add annotation to boolean output file + */ +process MergedRepConsensusPeakSetAnnotate { + label "process_medium" + publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: 'copy' + + when: + !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks + + input: + file bed from ch_mrep_macs_consensus_bed + file bool from ch_mrep_macs_consensus_bool + file fasta from ch_fasta + file gtf from ch_gtf + + output: + file "*.annotatePeaks.txt" into ch_mrep_macs_consensus_annotate + + script: + prefix = "consensus_peaks.mRp.clN" + """ + annotatePeaks.pl \\ + $bed \\ + $fasta \\ + -gid \\ + -gtf $gtf \\ + -cpu $task.cpus \\ + > ${prefix}.annotatePeaks.txt + + cut -f2- ${prefix}.annotatePeaks.txt | awk 'NR==1; NR > 1 {print \$0 | "sort -T '.' -k1,1 -k2,2n"}' | cut -f6- > tmp.txt + paste $bool tmp.txt > ${prefix}.boolean.annotatePeaks.txt + """ +} + +/* + * STEP 8.7 Count reads in consensus peaks with featureCounts and perform differential analysis with DESeq2 + */ +process MergedRepConsensusPeakSetDESeq { + label 'process_medium' + publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus/deseq2", mode: 'copy', + saveAs: { filename -> + if (filename.endsWith(".igv.txt")) null + else filename + } + + when: + !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks + + input: + file bams from ch_mlib_name_bam_mrep_counts.collect{ it[1] } + file saf from ch_mrep_macs_consensus_saf.collect() + file mrep_deseq2_pca_header from ch_mrep_deseq2_pca_header + file mrep_deseq2_clustering_header from ch_mrep_deseq2_clustering_header + + output: + file "*featureCounts.txt" into ch_mrep_macs_consensus_counts + file "*featureCounts.txt.summary" into ch_mrep_macs_consensus_counts_mqc + file "*.{RData,results.txt,pdf,log}" into ch_mrep_macs_consensus_deseq_results + file "sizeFactors" into ch_mrep_macs_consensus_deseq_factors + file "*vs*/*.{pdf,txt}" into ch_mrep_macs_consensus_deseq_comp_results + file "*vs*/*.bed" into ch_mrep_macs_consensus_deseq_comp_bed + file "*igv.txt" into ch_mrep_macs_consensus_deseq_comp_igv + file "*.tsv" into ch_mrep_macs_consensus_deseq_mqc + + script: + prefix = "consensus_peaks.mRp.clN" + bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() + bam_ext = params.single_end ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" + pe_params = params.single_end ? '' : "-p --donotsort" + """ + featureCounts \\ + -F SAF \\ + -O \\ + --fracOverlap 0.2 \\ + -T $task.cpus \\ + $pe_params \\ + -a $saf \\ + -o ${prefix}.featureCounts.txt \\ + ${bam_files.join(' ')} + + featurecounts_deseq2.r -i ${prefix}.featureCounts.txt -b '$bam_ext' -o ./ -p $prefix -s .mLb + + cat $mrep_deseq2_pca_header ${prefix}.pca.vals.txt > ${prefix}.pca.vals_mqc.tsv + cat $mrep_deseq2_clustering_header ${prefix}.sample.dists.txt > ${prefix}.sample.dists_mqc.tsv + + find * -type f -name "*.FDR0.05.results.bed" -exec echo -e "bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus/deseq2/"{}"\\t255,0,0" \\; > ${prefix}.igv.txt + """ +} + +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// +/* -- -- */ +/* -- IGV -- */ +/* -- -- */ +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// + +/* + * STEP 9 - Create IGV session file + */ +process IGV { + publishDir "${params.outdir}/igv/${PEAK_TYPE}", mode: 'copy' + + when: + !params.skip_igv + + input: + file fasta from ch_fasta + + file bigwigs from ch_mlib_bigwig_igv.collect().ifEmpty([]) + file peaks from ch_mlib_macs_igv.collect().ifEmpty([]) + file consensus_peaks from ch_mlib_macs_consensus_igv.collect().ifEmpty([]) + file differential_peaks from ch_mlib_macs_consensus_deseq_comp_igv.collect().ifEmpty([]) + + file rbigwigs from ch_mrep_bigwig_igv.collect().ifEmpty([]) + file rpeaks from ch_mrep_macs_igv.collect().ifEmpty([]) + file rconsensus_peaks from ch_mrep_macs_consensus_igv.collect().ifEmpty([]) + file rdifferential_peaks from ch_mrep_macs_consensus_deseq_comp_igv.collect().ifEmpty([]) + + output: + file "*.{txt,xml}" into ch_igv_session + + script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ + """ + cat *.txt > igv_files.txt + igv_files_to_session.py igv_session.xml igv_files.txt ../../reference_genome/${fasta.getName()} --path_prefix '../../' + """ +} + +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// +/* -- -- */ +/* -- MULTIQC -- */ +/* -- -- */ +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// + +/* + * Parse software version numbers + */ +process get_software_versions { + publishDir "${params.outdir}/pipeline_info", mode: 'copy', + saveAs: { filename -> + if (filename.indexOf(".csv") > 0) filename + else null + } + + output: + file 'software_versions_mqc.yaml' into ch_software_versions_mqc + file "software_versions.csv" + + script: + """ + echo $workflow.manifest.version > v_pipeline.txt + echo $workflow.nextflow.version > v_nextflow.txt + fastqc --version > v_fastqc.txt + trim_galore --version > v_trim_galore.txt + echo \$(bwa 2>&1) > v_bwa.txt + samtools --version > v_samtools.txt + bedtools --version > v_bedtools.txt + echo \$(bamtools --version 2>&1) > v_bamtools.txt + echo \$(plotFingerprint --version 2>&1) > v_deeptools.txt || true + picard MarkDuplicates --version &> v_picard.txt || true + echo \$(R --version 2>&1) > v_R.txt + python -c "import pysam; print(pysam.__version__)" > v_pysam.txt + echo \$(macs2 --version 2>&1) > v_macs2.txt + touch v_homer.txt + echo \$(ataqv --version 2>&1) > v_ataqv.txt + echo \$(featureCounts -v 2>&1) > v_featurecounts.txt + preseq &> v_preseq.txt + multiqc --version > v_multiqc.txt + scrape_software_versions.py &> software_versions_mqc.yaml + """ +} + +def create_workflow_summary(summary) { + + def yaml_file = workDir.resolve('workflow_summary_mqc.yaml') + yaml_file.text = """ + id: 'nf-core-atacseq-summary' + description: " - this information is collected when the pipeline is started." + section_name: 'nf-core/atacseq Workflow Summary' + section_href: 'https://github.com/nf-core/atacseq' + plot_type: 'html' + data: | +
+${summary.collect { k,v -> "
$k
${v ?: 'N/A'}
" }.join("\n")} +
+ """.stripIndent() + + return yaml_file +} + +/* + * STEP 10 - MultiQC + */ +process MultiQC { + publishDir "${params.outdir}/multiqc/${PEAK_TYPE}", mode: 'copy' + + when: + !params.skip_multiqc + + input: + file multiqc_config from ch_multiqc_config + + file ('software_versions/*') from ch_software_versions_mqc.collect() + file ('workflow_summary/*') from create_workflow_summary(summary) + + file ('fastqc/*') from ch_fastqc_reports_mqc.collect().ifEmpty([]) + file ('trimgalore/*') from ch_trimgalore_results_mqc.collect().ifEmpty([]) + file ('trimgalore/fastqc/*') from ch_trimgalore_fastqc_reports_mqc.collect().ifEmpty([]) + + file ('alignment/library/*') from ch_sort_bam_flagstat_mqc.collect() + + file ('alignment/mergedLibrary/*') from ch_mlib_bam_stats_mqc.collect() + file ('alignment/mergedLibrary/*') from ch_mlib_rm_orphan_flagstat_mqc.collect{it[1]} + file ('alignment/mergedLibrary/*') from ch_mlib_rm_orphan_stats_mqc.collect() + file ('alignment/mergedLibrary/picard_metrics/*') from ch_mlib_bam_metrics_mqc.collect() + file ('alignment/mergedLibrary/picard_metrics/*') from ch_mlib_collectmetrics_mqc.collect() + file ('macs/mergedLibrary/*') from ch_mlib_macs_mqc.collect().ifEmpty([]) + file ('macs/mergedLibrary/*') from ch_mlib_peak_qc_mqc.collect().ifEmpty([]) + file ('macs/mergedLibrary/consensus/*') from ch_mlib_macs_consensus_counts_mqc.collect().ifEmpty([]) + file ('macs/mergedLibrary/consensus/*') from ch_mlib_macs_consensus_deseq_mqc.collect().ifEmpty([]) + file ('preseq/*') from ch_mlib_preseq_mqc.collect().ifEmpty([]) + file ('deeptools/*') from ch_mlib_plotprofile_mqc.collect().ifEmpty([]) + file ('deeptools/*') from ch_mlib_plotfingerprint_mqc.collect().ifEmpty([]) + + file ('alignment/mergedReplicate/*') from ch_mrep_bam_flagstat_mqc.collect{it[1]}.ifEmpty([]) + file ('alignment/mergedReplicate/*') from ch_mrep_bam_stats_mqc.collect().ifEmpty([]) + file ('alignment/mergedReplicate/*') from ch_mrep_bam_metrics_mqc.collect().ifEmpty([]) + file ('macs/mergedReplicate/*') from ch_mrep_macs_mqc.collect().ifEmpty([]) + file ('macs/mergedReplicate/*') from ch_mrep_peak_qc_mqc.collect().ifEmpty([]) + file ('macs/mergedReplicate/consensus/*') from ch_mrep_macs_consensus_counts_mqc.collect().ifEmpty([]) + file ('macs/mergedReplicate/consensus/*') from ch_mrep_macs_consensus_deseq_mqc.collect().ifEmpty([]) + + output: + file "*multiqc_report.html" into ch_multiqc_report + file "*_data" + file "multiqc_plots" + + script: + rtitle = custom_runName ? "--title \"$custom_runName\"" : '' + rfilename = custom_runName ? "--filename " + custom_runName.replaceAll('\\W','_').replaceAll('_+','_') + "_multiqc_report" : '' + """ + multiqc . -f $rtitle $rfilename --config $multiqc_config \\ + -m custom_content -m fastqc -m cutadapt -m samtools -m picard -m preseq -m featureCounts -m deeptools + """ +} + +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// +/* -- -- */ +/* -- REPORTS/DOCUMENTATION -- */ +/* -- -- */ +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// + +/* + * STEP 11 - Output description HTML + */ +process output_documentation { + publishDir "${params.outdir}/Documentation", mode: 'copy' + + input: + file output_docs from ch_output_docs + + output: + file "results_description.html" + + script: + """ + markdown_to_html.r $output_docs results_description.html + """ +} + +/* + * Completion e-mail notification + */ +workflow.onComplete { + + // Set up the e-mail variables + def subject = "[nf-core/atacseq] Successful: $workflow.runName" + if (!workflow.success) { + subject = "[nf-core/atacseq] FAILED: $workflow.runName" + } + def email_fields = [:] + email_fields['version'] = workflow.manifest.version + email_fields['runName'] = custom_runName ?: workflow.runName + email_fields['success'] = workflow.success + email_fields['dateComplete'] = workflow.complete + email_fields['duration'] = workflow.duration + email_fields['exitStatus'] = workflow.exitStatus + email_fields['errorMessage'] = (workflow.errorMessage ?: 'None') + email_fields['errorReport'] = (workflow.errorReport ?: 'None') + email_fields['commandLine'] = workflow.commandLine + email_fields['projectDir'] = workflow.projectDir + email_fields['summary'] = summary + email_fields['summary']['Date Started'] = workflow.start + email_fields['summary']['Date Completed'] = workflow.complete + email_fields['summary']['Pipeline script file path'] = workflow.scriptFile + email_fields['summary']['Pipeline script hash ID'] = workflow.scriptId + if (workflow.repository) email_fields['summary']['Pipeline repository Git URL'] = workflow.repository + if (workflow.commitId) email_fields['summary']['Pipeline repository Git Commit'] = workflow.commitId + if (workflow.revision) email_fields['summary']['Pipeline Git branch/tag'] = workflow.revision + if (workflow.container) email_fields['summary']['Docker image'] = workflow.container + email_fields['summary']['Nextflow Version'] = workflow.nextflow.version + email_fields['summary']['Nextflow Build'] = workflow.nextflow.build + email_fields['summary']['Nextflow Compile Timestamp'] = workflow.nextflow.timestamp + + // On success try attach the multiqc report + def mqc_report = null + try { + if (workflow.success) { + mqc_report = ch_multiqc_report.getVal() + if (mqc_report.getClass() == ArrayList) { + log.warn "[nf-core/atacseq] Found multiple reports from process 'multiqc', will use only one" + mqc_report = mqc_report[0] + } + } + } catch (all) { + log.warn "[nf-core/atacseq] Could not attach MultiQC report to summary email" + } + + // Check if we are only sending emails on failure + email_address = params.email + if (!params.email && params.email_on_fail && !workflow.success) { + email_address = params.email_on_fail + } + + // Render the TXT template + def engine = new groovy.text.GStringTemplateEngine() + def tf = new File("$baseDir/assets/email_template.txt") + def txt_template = engine.createTemplate(tf).make(email_fields) + def email_txt = txt_template.toString() + + // Render the HTML template + def hf = new File("$baseDir/assets/email_template.html") + def html_template = engine.createTemplate(hf).make(email_fields) + def email_html = html_template.toString() + + // Render the sendmail template + def smail_fields = [ email: email_address, subject: subject, email_txt: email_txt, email_html: email_html, baseDir: "$baseDir", mqcFile: mqc_report, mqcMaxSize: params.max_multiqc_email_size.toBytes() ] + def sf = new File("$baseDir/assets/sendmail_template.txt") + def sendmail_template = engine.createTemplate(sf).make(smail_fields) + def sendmail_html = sendmail_template.toString() + + // Send the HTML e-mail + if (email_address) { + try { + if (params.plaintext_email) { throw GroovyException('Send plaintext e-mail, not HTML') } + // Try to send HTML e-mail using sendmail + [ 'sendmail', '-t' ].execute() << sendmail_html + log.info "[nf-core/atacseq] Sent summary e-mail to $email_address (sendmail)" + } catch (all) { + // Catch failures and try with plaintext + [ 'mail', '-s', subject, email_address ].execute() << email_txt + log.info "[nf-core/atacseq] Sent summary e-mail to $email_address (mail)" + } + } + + // Write summary e-mail HTML to a file + def output_d = new File("${params.outdir}/pipeline_info/") + if (!output_d.exists()) { + output_d.mkdirs() + } + def output_hf = new File(output_d, "pipeline_report.html") + output_hf.withWriter { w -> w << email_html } + def output_tf = new File(output_d, "pipeline_report.txt") + output_tf.withWriter { w -> w << email_txt } + + c_reset = params.monochrome_logs ? '' : "\033[0m"; + c_purple = params.monochrome_logs ? '' : "\033[0;35m"; + c_green = params.monochrome_logs ? '' : "\033[0;32m"; + c_red = params.monochrome_logs ? '' : "\033[0;31m"; + + if (workflow.stats.ignoredCount > 0 && workflow.success) { + log.info "${c_purple}Warning, pipeline completed, but with errored process(es) ${c_reset}" + log.info "${c_red}Number of ignored errored process(es) : ${workflow.stats.ignoredCount} ${c_reset}" + log.info "${c_green}Number of successfully ran process(es) : ${workflow.stats.succeedCount} ${c_reset}" + } + + if (workflow.success) { + log.info "${c_purple}[nf-core/atacseq]${c_green} Pipeline completed successfully${c_reset}" + } else { + checkHostname() + log.info "${c_purple}[nf-core/atacseq]${c_red} Pipeline completed with errors${c_reset}" + } + +} /////////////////////////////////////////////////////////////////////////////// /////////////////////////////////////////////////////////////////////////////// From 70b6c32a1485e36a533966b04ae7bad41961eabd Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 31 Jan 2020 09:38:27 +0000 Subject: [PATCH 082/226] Update CHANGELOG --- CHANGELOG.md | 1 + 1 file changed, 1 insertion(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index f727f28c..d533e46a 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -11,6 +11,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * [#63](https://github.com/nf-core/atacseq/issues/63) - Added multicore support for Trim Galore! * [#75](https://github.com/nf-core/atacseq/issues/75) - Include gene annotation versions in multiqc report +* [#76](https://github.com/nf-core/atacseq/issues/76) - featureCounts coupled to DESeq2 * Added GitHub Actions CI tests ### `Fixed` From aab71886d01e09d52a3707bd97063871cb2cefd2 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 31 Jan 2020 09:38:43 +0000 Subject: [PATCH 083/226] Add featureCounts in separate process --- main.nf | 114 ++++++++++++++++++++++++++++++++++++++------------------ 1 file changed, 77 insertions(+), 37 deletions(-) diff --git a/main.nf b/main.nf index e6294f73..00072f6f 100644 --- a/main.nf +++ b/main.nf @@ -1275,39 +1275,26 @@ process MergedLibConsensusPeakSetAnnotate { } /* - * STEP 6.6 Count reads in consensus peaks with featureCounts and perform differential analysis with DESeq2 + * STEP 6.6 Count reads in consensus peaks with featureCounts */ -process MergedLibConsensusPeakSetDESeq { +process MergedLibConsensusPeakSetCounts { label 'process_medium' - publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus/deseq2", mode: 'copy', - saveAs: { filename -> - if (filename.endsWith(".igv.txt")) null - else filename - } + publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: 'copy' when: - params.macs_gsize && replicatesExist && multipleGroups && !params.skip_consensus_peaks + params.macs_gsize && (replicatesExist || multipleGroups) && !params.skip_consensus_peaks input: file bams from ch_mlib_name_bam_mlib_counts.collect{ it[1] } file saf from ch_mlib_macs_consensus_saf.collect() - file mlib_deseq2_pca_header from ch_mlib_deseq2_pca_header - file mlib_deseq2_clustering_header from ch_mlib_deseq2_clustering_header output: file "*featureCounts.txt" into ch_mlib_macs_consensus_counts file "*featureCounts.txt.summary" into ch_mlib_macs_consensus_counts_mqc - file "*.{RData,results.txt,pdf,log}" into ch_mlib_macs_consensus_deseq_results - file "sizeFactors" into ch_mlib_macs_consensus_deseq_factors - file "*vs*/*.{pdf,txt}" into ch_mlib_macs_consensus_deseq_comp_results - file "*vs*/*.bed" into ch_mlib_macs_consensus_deseq_comp_bed - file "*igv.txt" into ch_mlib_macs_consensus_deseq_comp_igv - file "*.tsv" into ch_mlib_macs_consensus_deseq_mqc script: prefix = "consensus_peaks.mLb.clN" bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() - bam_ext = params.single_end ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" pe_params = params.single_end ? '' : "-p --donotsort" """ featureCounts \\ @@ -1319,8 +1306,41 @@ process MergedLibConsensusPeakSetDESeq { -a $saf \\ -o ${prefix}.featureCounts.txt \\ ${bam_files.join(' ')} + """ +} + +/* + * STEP 6.7 Perform differential analysis with DESeq2 + */ +process MergedLibConsensusPeakSetDESeq { + label 'process_medium' + publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus/deseq2", mode: 'copy', + saveAs: { filename -> + if (filename.endsWith(".igv.txt")) null + else filename + } + + when: + params.macs_gsize && replicatesExist && multipleGroups && !params.skip_consensus_peaks + + input: + file counts from ch_mlib_macs_consensus_counts + file mlib_deseq2_pca_header from ch_mlib_deseq2_pca_header + file mlib_deseq2_clustering_header from ch_mlib_deseq2_clustering_header + + output: + file "*.{RData,results.txt,pdf,log}" into ch_mlib_macs_consensus_deseq_results + file "sizeFactors" into ch_mlib_macs_consensus_deseq_factors + file "*vs*/*.{pdf,txt}" into ch_mlib_macs_consensus_deseq_comp_results + file "*vs*/*.bed" into ch_mlib_macs_consensus_deseq_comp_bed + file "*igv.txt" into ch_mlib_macs_consensus_deseq_comp_igv + file "*.tsv" into ch_mlib_macs_consensus_deseq_mqc - featurecounts_deseq2.r -i ${prefix}.featureCounts.txt -b '$bam_ext' -o ./ -p $prefix -s .mLb + script: + prefix = "consensus_peaks.mLb.clN" + bam_ext = params.single_end ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" + """ + featurecounts_deseq2.r -i $counts -b '$bam_ext' -o ./ -p $prefix -s .mLb cat $mlib_deseq2_pca_header ${prefix}.pca.vals.txt > ${prefix}.pca.vals_mqc.tsv cat $mlib_deseq2_clustering_header ${prefix}.sample.dists.txt > ${prefix}.sample.dists_mqc.tsv @@ -1330,7 +1350,7 @@ process MergedLibConsensusPeakSetDESeq { } /* - * STEP 6.7 Run ataqv on BAM file and corresponding peaks + * STEP 6.8 Run ataqv on BAM file and corresponding peaks */ process MergedLibAtaqv { tag "$name" @@ -1367,7 +1387,7 @@ process MergedLibAtaqv { } /* - * STEP 6.8 run ataqv mkarv on all JSON files to render web app + * STEP 6.9 run ataqv mkarv on all JSON files to render web app */ process MergedLibAtaqvMkarv { label 'process_medium' @@ -1741,15 +1761,11 @@ process MergedRepConsensusPeakSetAnnotate { } /* - * STEP 8.7 Count reads in consensus peaks with featureCounts and perform differential analysis with DESeq2 + * STEP 8.7 Count reads in consensus peaks with featureCounts */ -process MergedRepConsensusPeakSetDESeq { +process MergedRepConsensusPeakSetCounts { label 'process_medium' - publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus/deseq2", mode: 'copy', - saveAs: { filename -> - if (filename.endsWith(".igv.txt")) null - else filename - } + publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: 'copy' when: !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks @@ -1757,23 +1773,14 @@ process MergedRepConsensusPeakSetDESeq { input: file bams from ch_mlib_name_bam_mrep_counts.collect{ it[1] } file saf from ch_mrep_macs_consensus_saf.collect() - file mrep_deseq2_pca_header from ch_mrep_deseq2_pca_header - file mrep_deseq2_clustering_header from ch_mrep_deseq2_clustering_header output: file "*featureCounts.txt" into ch_mrep_macs_consensus_counts file "*featureCounts.txt.summary" into ch_mrep_macs_consensus_counts_mqc - file "*.{RData,results.txt,pdf,log}" into ch_mrep_macs_consensus_deseq_results - file "sizeFactors" into ch_mrep_macs_consensus_deseq_factors - file "*vs*/*.{pdf,txt}" into ch_mrep_macs_consensus_deseq_comp_results - file "*vs*/*.bed" into ch_mrep_macs_consensus_deseq_comp_bed - file "*igv.txt" into ch_mrep_macs_consensus_deseq_comp_igv - file "*.tsv" into ch_mrep_macs_consensus_deseq_mqc script: prefix = "consensus_peaks.mRp.clN" bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() - bam_ext = params.single_end ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" pe_params = params.single_end ? '' : "-p --donotsort" """ featureCounts \\ @@ -1785,8 +1792,41 @@ process MergedRepConsensusPeakSetDESeq { -a $saf \\ -o ${prefix}.featureCounts.txt \\ ${bam_files.join(' ')} + """ +} + +/* + * STEP 8.8 Perform differential analysis with DESeq2 + */ +process MergedRepConsensusPeakSetDESeq { + label 'process_medium' + publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus/deseq2", mode: 'copy', + saveAs: { filename -> + if (filename.endsWith(".igv.txt")) null + else filename + } + + when: + !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks + + input: + file counts from ch_mrep_macs_consensus_counts + file mrep_deseq2_pca_header from ch_mrep_deseq2_pca_header + file mrep_deseq2_clustering_header from ch_mrep_deseq2_clustering_header - featurecounts_deseq2.r -i ${prefix}.featureCounts.txt -b '$bam_ext' -o ./ -p $prefix -s .mLb + output: + file "*.{RData,results.txt,pdf,log}" into ch_mrep_macs_consensus_deseq_results + file "sizeFactors" into ch_mrep_macs_consensus_deseq_factors + file "*vs*/*.{pdf,txt}" into ch_mrep_macs_consensus_deseq_comp_results + file "*vs*/*.bed" into ch_mrep_macs_consensus_deseq_comp_bed + file "*igv.txt" into ch_mrep_macs_consensus_deseq_comp_igv + file "*.tsv" into ch_mrep_macs_consensus_deseq_mqc + + script: + prefix = "consensus_peaks.mRp.clN" + bam_ext = params.single_end ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" + """ + featurecounts_deseq2.r -i $counts -b '$bam_ext' -o ./ -p $prefix -s .mLb cat $mrep_deseq2_pca_header ${prefix}.pca.vals.txt > ${prefix}.pca.vals_mqc.tsv cat $mrep_deseq2_clustering_header ${prefix}.sample.dists.txt > ${prefix}.sample.dists_mqc.tsv From a46663ea354668562464b29dadaecd3ea9462315 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 31 Jan 2020 10:04:32 +0000 Subject: [PATCH 084/226] Remove Travis badge --- README.md | 1 - 1 file changed, 1 deletion(-) diff --git a/README.md b/README.md index eab86f7f..c40fc903 100644 --- a/README.md +++ b/README.md @@ -1,6 +1,5 @@ # ![nfcore/atacseq](docs/images/nf-core-atacseq_logo.png) -[![Build Status](https://travis-ci.com/nf-core/atacseq.svg?branch=master)](https://travis-ci.com/nf-core/atacseq) [![GitHub Actions CI Status](https://github.com/nf-core/atacseq/workflows/nf-core%20CI/badge.svg)](https://github.com/nf-core/atacseq/actions) [![GitHub Actions Linting Status](https://github.com/nf-core/atacseq/workflows/nf-core%20linting/badge.svg)](https://github.com/nf-core/atacseq/actions) [![Nextflow](https://img.shields.io/badge/nextflow-%E2%89%A519.10.0-brightgreen.svg)](https://www.nextflow.io/) From ccb2175141906cf8fa7660858b41dbf5b418c346 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 31 Jan 2020 10:15:16 +0000 Subject: [PATCH 085/226] Update output docs --- docs/output.md | 16 ++++++++-------- 1 file changed, 8 insertions(+), 8 deletions(-) diff --git a/docs/output.md b/docs/output.md index 6ad1b844..aef77550 100755 --- a/docs/output.md +++ b/docs/output.md @@ -172,10 +172,10 @@ The library-level alignments associated with the same sample are merged and subs * QC plots for peak-to-gene feature annotation: `macs_annotatePeaks.plots.pdf` * MultiQC custom-content files for FRiP score, peak count and peak-to-gene ratios: `*.FRiP_mqc.tsv`, `*.count_mqc.tsv` and `macs_annotatePeaks.summary_mqc.tsv` respectively. -5. **Create consensus set of peaks** +5. **Create and quantify consensus set of peaks** *Documentation*: - [BEDTools](https://bedtools.readthedocs.io/en/latest/) + [BEDTools](https://bedtools.readthedocs.io/en/latest/), [featureCounts](http://bioinf.wehi.edu.au/featureCounts/) *Description*: In order to perform the differential binding analysis we need to be able to carry out the read quantification for the same intervals across **all** of the samples in the experiment. To this end, the individual peak-sets called per sample have to be merged together in order to create a consensus set of peaks. @@ -186,10 +186,15 @@ The library-level alignments associated with the same sample are merged and subs ![R - UpSetR peak intersection plot](images/r_upsetr_intersect_plot.png) + The featureCounts tool is used to count the number of reads relative to the consensus peak-set across all of the samples. This essentially generates a file containing a matrix where the rows represent the consensus intervals, the columns represent all of the samples in the experiment, and the values represent the raw read counts. + + ![MultiQC - featureCounts consensus peak read assignment plot](images/mqc_featureCounts_assignment_plot.png) + *Output directories*: * `bwa/mergedLibrary/macs//consensus/` * Consensus peak-set across all samples in `*.bed` format. * Consensus peak-set across all samples in `*.saf` format. Required by featureCounts for read quantification. + * `.featureCounts.txt` file for read counts across all samples relative to consensus peak-set. * HOMER `*.annotatePeaks.txt` peak-to-gene annotation file for consensus peaks. * Spreadsheet representation of consensus peak-set across samples **with** gene annotation columns: `*.boolean.annotatePeaks.txt`. The columns from individual peak files are included in this file along with the ability to filter peaks based on their presence or absence in multiple replicates/conditions. @@ -200,13 +205,9 @@ The library-level alignments associated with the same sample are merged and subs 6. **Read counting and differential binding analysis** *Documentation*: - [featureCounts](http://bioinf.wehi.edu.au/featureCounts/), [DESeq2](https://bioconductor.org/packages/release/bioc/vignettes/DESeq2/inst/doc/DESeq2.html), [R](https://www.r-project.org/) + [DESeq2](https://bioconductor.org/packages/release/bioc/vignettes/DESeq2/inst/doc/DESeq2.html), [R](https://www.r-project.org/) *Description*: - The featureCounts tool is used to count the number of reads relative to the consensus peak-set across all of the samples. This essentially generates a file containing a matrix where the rows represent the consensus intervals, the columns represent all of the samples in the experiment, and the values represent the raw read counts. - - ![MultiQC - featureCounts consensus peak read assignment plot](images/mqc_featureCounts_assignment_plot.png) - DESeq2 is more commonly used to perform differential expression analysis for RNA-seq datasets. However, it can also be used for ATAC-seq differential accessibility analysis, in which case you can imagine that instead of counts per gene for RNA-seq data we now have counts per accessible region. This pipeline uses a standardised DESeq2 analysis script to get an idea of the reproducibility within the experiment, and to assess the overall differential binding. Please note that this will not suit every experimental design, and if there are other problems with the experiment then it may not work as well as expected. @@ -223,7 +224,6 @@ The library-level alignments associated with the same sample are merged and subs *Output directories*: * `bwa/mergedLibrary/macs//consensus//deseq2/` - * `.featureCounts.txt` file for read counts across all samples relative to consensus peak-set. * Differential binding `*.results.txt` spreadsheet containing results across all consensus peaks and all comparisons. * `*.plots.pdf` file for PCA and hierarchical clustering. * `*.log` file with information for number of differentially bound intervals at different FDR and fold-change thresholds for each comparison. From 2a7f66a295eca7ced6fc3f43aa4574650639a9e6 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 31 Jan 2020 17:41:57 +0000 Subject: [PATCH 086/226] Fix Docker --- Dockerfile | 0 1 file changed, 0 insertions(+), 0 deletions(-) mode change 100755 => 100644 Dockerfile diff --git a/Dockerfile b/Dockerfile old mode 100755 new mode 100644 From 74f3de7936c5e902ee5a37bfe555c413bdfe239f Mon Sep 17 00:00:00 2001 From: MaxUlysse Date: Tue, 11 Feb 2020 14:48:35 +0100 Subject: [PATCH 087/226] add social preview image rename files --- assets/nf-core-atacseq_social_preview.png | Bin 0 -> 51685 bytes assets/nf-core-atacseq_social_preview.svg | 448 ++++++++++++++++++++++ 2 files changed, 448 insertions(+) create mode 100644 assets/nf-core-atacseq_social_preview.png create mode 100644 assets/nf-core-atacseq_social_preview.svg diff --git a/assets/nf-core-atacseq_social_preview.png b/assets/nf-core-atacseq_social_preview.png new file mode 100644 index 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z_bez2%w&zEv9M4_z?G>bpi@!NZvT(XY9_1YW7y2~F;a+&cGvGF*lrP7=RMjS@`;P<8rk)~FdARr&Z|MKa z0Iu17GN%!#RUH#IT};dfddfd;*|G%!`98+!z3BkNq9ZRwQQYs<)_zxoZ(y7b)nNoC z3w1cC=b1)1s!+AIb=%KPzT&lTC6~I*Bc#~D;^KMN-W~RQT{Tx9%EBo5IqS;N>HXPi zBTuOJ=jA`j*YNTSd3hLK-j0_S;bm0(zo&`QJ^cb&ca|2%IUbal + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + image/svg+xml + + + + + + + ATAC-seq peak-calling, QC and differential analysis pipeline + atacseq + + + + + + + + + + + + + + + + + + + + + + + + + From 766a1f375e49a8a9db5b0004229d10776c02b141 Mon Sep 17 00:00:00 2001 From: MaxUlysse Date: Tue, 11 Feb 2020 15:41:50 +0100 Subject: [PATCH 088/226] update CHANGELOG --- CHANGELOG.md | 1 + 1 file changed, 1 insertion(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index d533e46a..7c54f2f1 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -12,6 +12,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * [#63](https://github.com/nf-core/atacseq/issues/63) - Added multicore support for Trim Galore! * [#75](https://github.com/nf-core/atacseq/issues/75) - Include gene annotation versions in multiqc report * [#76](https://github.com/nf-core/atacseq/issues/76) - featureCounts coupled to DESeq2 +* [#80](https://github.com/nf-core/atacseq/pull/80) - Added social preview image * Added GitHub Actions CI tests ### `Fixed` From 7c2956adb1aeb196ef3daba711f8596081d4d0ea Mon Sep 17 00:00:00 2001 From: drpatelh Date: Mon, 17 Feb 2020 17:45:55 +0000 Subject: [PATCH 089/226] Update packages --- CHANGELOG.md | 17 ++++++++++------- environment.yml | 21 ++++++++++----------- 2 files changed, 20 insertions(+), 18 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 7c54f2f1..e03ca31f 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -24,26 +24,29 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Dependencies` -* Add genrich `0.6` * Add r-stringi `1.4.3` * Add pigz `2.3.4` * Update gawk `4.2.1` -> `5.0.1` -* Update r-base `3.4.1` -> `3.6.1` +* Update r-base `3.4.1` -> `3.6.2` * Update r-optparse `1.6.0` -> `1.6.4` * Update r-ggplot2 `3.1.0` -> `3.2.1` * Update r-pheatmap `1.0.10` -> `1.0.12` * Update r-lattice `0.20_35` -> `0.20_38` * Update r-upsetr `1.3.3` -> `1.4.0` * Update r-scales `1.0.0` -> `1.1.0` -* Update r-xfun `0.3` -> `0.11` +* Update r-xfun `0.3` -> `0.12` +* Update fastqc `0.11.8` -> `0.11.9` * Update trim-galore `0.5.0` -> `0.6.5` -* Update picard `2.19.0` -> `2.21.3` +* Update samtools `1.9` -> `1.10` +* Update picard `2.19.0` -> `2.21.8` * Update pysam `0.15.2` -> `0.15.3` -* Update bedtools `2.27.1` -> `2.29.0` +* Update bedtools `2.27.1` -> `2.29.2` * Update ucsc-bedgraphtobigwig `377` -> `357` -* Update deeptools `3.2.1` -> `3.3.1` -* Update macs2 `2.1.2` -> `2.2.5` +* Update deeptools `3.2.1` -> `3.3.2` +* Update macs2 `2.1.2` -> `2.2.6` * Update homer `4.9.1` -> `4.10` +* Update ataqv `1.0.0` -> `1.1.1` +* Update subread `1.6.4` -> `2.0.0` * Update multiqc `1.7` -> `1.8` * Update bioconductor-deseq2 `1.20.0` -> `1.26.0` * Update bioconductor-vsn `3.46.0` -> `3.54.0` diff --git a/environment.yml b/environment.yml index b89ccc24..2fc1a29d 100755 --- a/environment.yml +++ b/environment.yml @@ -8,7 +8,7 @@ channels: dependencies: ## conda-forge packages - conda-forge::gawk=5.0.1 - - conda-forge::r-base=3.6.1 + - conda-forge::r-base=3.6.2 - conda-forge::r-optparse=1.6.4 - conda-forge::r-rcolorbrewer=1.1_2 - conda-forge::r-ggplot2=3.2.1 @@ -17,26 +17,25 @@ dependencies: - conda-forge::r-pheatmap=1.0.12 - conda-forge::r-lattice=0.20_38 - conda-forge::r-upsetr=1.4.0 - - conda-forge::r-xfun=0.11 + - conda-forge::r-xfun=0.12 - conda-forge::pigz=2.3.4 - r::r-stringi=1.4.3 ## bioconda packages - - bioconda::fastqc=0.11.8 + - bioconda::fastqc=0.11.9 - bioconda::trim-galore=0.6.5 - bioconda::bwa=0.7.17 - - bioconda::samtools=1.9 - - bioconda::picard=2.21.3 + - bioconda::samtools=1.10 + - bioconda::picard=2.21.8 - bioconda::bamtools=2.5.1 - bioconda::pysam=0.15.3 - - bioconda::bedtools=2.29.0 + - bioconda::bedtools=2.29.2 - bioconda::ucsc-bedgraphtobigwig=357 - - bioconda::deeptools=3.3.1 - - bioconda::macs2=2.2.5 - - bioconda::genrich=0.6 + - bioconda::deeptools=3.3.2 + - bioconda::macs2=2.2.6 - bioconda::homer=4.10 - - bioconda::ataqv=1.0.0 - - bioconda::subread=1.6.4 + - bioconda::ataqv=1.1.1 + - bioconda::subread=2.0.0 - bioconda::preseq=2.0.3 - bioconda::multiqc=1.8 - bioconda::bioconductor-deseq2=1.26.0 From 7d4e7833f86882e42a976577ab43ea354195d750 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Mon, 17 Feb 2020 17:55:14 +0000 Subject: [PATCH 090/226] Downgrade samtools --- CHANGELOG.md | 3 +-- environment.yml | 4 ++-- 2 files changed, 3 insertions(+), 4 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index e03ca31f..35497d86 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -37,9 +37,8 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * Update r-xfun `0.3` -> `0.12` * Update fastqc `0.11.8` -> `0.11.9` * Update trim-galore `0.5.0` -> `0.6.5` -* Update samtools `1.9` -> `1.10` * Update picard `2.19.0` -> `2.21.8` -* Update pysam `0.15.2` -> `0.15.3` +* Update pysam `0.15.2` -> `0.15.4` * Update bedtools `2.27.1` -> `2.29.2` * Update ucsc-bedgraphtobigwig `377` -> `357` * Update deeptools `3.2.1` -> `3.3.2` diff --git a/environment.yml b/environment.yml index 2fc1a29d..7381aa34 100755 --- a/environment.yml +++ b/environment.yml @@ -25,10 +25,10 @@ dependencies: - bioconda::fastqc=0.11.9 - bioconda::trim-galore=0.6.5 - bioconda::bwa=0.7.17 - - bioconda::samtools=1.10 + - bioconda::samtools=1.9 - bioconda::picard=2.21.8 - bioconda::bamtools=2.5.1 - - bioconda::pysam=0.15.3 + - bioconda::pysam=0.15.4 - bioconda::bedtools=2.29.2 - bioconda::ucsc-bedgraphtobigwig=357 - bioconda::deeptools=3.3.2 From 04a429f6016741a1e31b83147f384315318d1191 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 19 Feb 2020 14:08:45 +0000 Subject: [PATCH 091/226] Update with template --- .github/workflows/ci.yml | 16 +++++++++------- 1 file changed, 9 insertions(+), 7 deletions(-) diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml index fa664e06..a442aa32 100755 --- a/.github/workflows/ci.yml +++ b/.github/workflows/ci.yml @@ -5,22 +5,24 @@ on: [push, pull_request] jobs: test: - runs-on: ubuntu-18.04 + env: + NXF_VER: ${{ matrix.nxf_ver }} + NXF_ANSI_LOG: false + runs-on: ubuntu-latest strategy: matrix: # Nextflow versions: check pipeline minimum and current latest nxf_ver: ['19.10.0', ''] steps: - - uses: actions/checkout@v1 + - uses: actions/checkout@v2 - name: Install Nextflow run: | - export NXF_VER=${{ matrix.nxf_ver }} - export NXF_ANSI_LOG=false wget -qO- get.nextflow.io | bash sudo mv nextflow /usr/local/bin/ - - name: Pull image + - name: Pull docker image run: | - docker pull nfcore/atacseq:dev && docker tag nfcore/atacseq:dev nfcore/atacseq:dev - - name: Run test + docker pull nfcore/atacseq:dev + docker tag nfcore/atacseq:dev nfcore/atacseq:dev + - name: Run pipeline with test data run: | nextflow run ${GITHUB_WORKSPACE} -profile test,docker From d8eab7705afc349efd740416ee0b82db83ed3bc7 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 19 Feb 2020 15:19:04 +0000 Subject: [PATCH 092/226] Update packages in environment --- CHANGELOG.md | 4 +++- environment.yml | 6 +++--- main.nf | 2 +- 3 files changed, 7 insertions(+), 5 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 35497d86..1c52bb6e 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -24,8 +24,9 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Dependencies` -* Add r-stringi `1.4.3` * Add pigz `2.3.4` +* Add r-tidyr `1.0.2` +* Add bioconductor-biocparallel `1.20.0` * Update gawk `4.2.1` -> `5.0.1` * Update r-base `3.4.1` -> `3.6.2` * Update r-optparse `1.6.0` -> `1.6.4` @@ -49,6 +50,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * Update multiqc `1.7` -> `1.8` * Update bioconductor-deseq2 `1.20.0` -> `1.26.0` * Update bioconductor-vsn `3.46.0` -> `3.54.0` +* Remove r-reshape2 `1.4.3` ### `Deprecated` diff --git a/environment.yml b/environment.yml index 7381aa34..e826b8be 100755 --- a/environment.yml +++ b/environment.yml @@ -12,14 +12,13 @@ dependencies: - conda-forge::r-optparse=1.6.4 - conda-forge::r-rcolorbrewer=1.1_2 - conda-forge::r-ggplot2=3.2.1 - - conda-forge::r-reshape2=1.4.3 + - conda-forge::r-tidyr=1.0.2 - conda-forge::r-scales=1.1.0 - conda-forge::r-pheatmap=1.0.12 - conda-forge::r-lattice=0.20_38 - conda-forge::r-upsetr=1.4.0 - conda-forge::r-xfun=0.12 - - conda-forge::pigz=2.3.4 - - r::r-stringi=1.4.3 + - conda-forge::pigz=2.3.4 ## Required for TrimGalore multi-threading ## bioconda packages - bioconda::fastqc=0.11.9 @@ -38,5 +37,6 @@ dependencies: - bioconda::subread=2.0.0 - bioconda::preseq=2.0.3 - bioconda::multiqc=1.8 + - bioconda::bioconductor-biocparallel=1.20.0 - bioconda::bioconductor-deseq2=1.26.0 - bioconda::bioconductor-vsn=3.54.0 diff --git a/main.nf b/main.nf index 00072f6f..6a384e1b 100644 --- a/main.nf +++ b/main.nf @@ -450,8 +450,8 @@ process MakeGenomeFilter { output: file "$fasta" // FASTA FILE FOR IGV - file "$readme" // AWS IGENOMES FILE CONTAINING ANNOTATION VERSION file "*.fai" // FAI INDEX FOR REFERENCE GENOME + //file "$readme" // AWS IGENOMES FILE CONTAINING ANNOTATION VERSION file "*.bed" into ch_genome_filter_regions // BED FILE WITHOUT BLACKLIST REGIONS & MITOCHONDRIAL CONTIG FOR FILTERING file "*.txt" into ch_genome_autosomes // TEXT FILE CONTAINING LISTING OF AUTOSOMAL CHROMOSOMES FOR ATAQV file "*.sizes" into ch_genome_sizes_mlib_bigwig, // CHROMOSOME SIZES FILE FOR BEDTOOLS From e7beca5123bcd35e91b3a66a445cf7ab4402d224 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 19 Feb 2020 16:27:51 +0000 Subject: [PATCH 093/226] Fix linting --- .github/workflows/ci.yml | 3 +-- 1 file changed, 1 insertion(+), 2 deletions(-) diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml index a442aa32..0fd5b152 100755 --- a/.github/workflows/ci.yml +++ b/.github/workflows/ci.yml @@ -21,8 +21,7 @@ jobs: sudo mv nextflow /usr/local/bin/ - name: Pull docker image run: | - docker pull nfcore/atacseq:dev - docker tag nfcore/atacseq:dev nfcore/atacseq:dev + docker pull nfcore/atacseq:dev && docker tag nfcore/atacseq:dev nfcore/atacseq:dev - name: Run pipeline with test data run: | nextflow run ${GITHUB_WORKSPACE} -profile test,docker From e6c7d97dc712581e3759bbad65a74a38b047ed5e Mon Sep 17 00:00:00 2001 From: runner Date: Thu, 20 Feb 2020 15:29:38 +0000 Subject: [PATCH 094/226] Template update for nf-core/tools version 1.9 --- .github/CONTRIBUTING.md | 52 ++- .github/ISSUE_TEMPLATE/bug_report.md | 43 ++- .github/ISSUE_TEMPLATE/feature_request.md | 18 +- .github/PULL_REQUEST_TEMPLATE.md | 28 +- .github/markdownlint.yml | 4 - .github/workflows/branch.yml | 16 + .github/workflows/ci.yml | 30 ++ .github/workflows/linting.yml | 50 +++ .gitignore | 3 +- .travis.yml | 42 -- CHANGELOG.md | 14 +- CODE_OF_CONDUCT.md | 2 +- Dockerfile | 12 +- README.md | 53 ++- assets/email_template.html | 2 + assets/email_template.txt | 12 +- assets/multiqc_config.yaml | 4 +- assets/nf-core-atacseq_logo.png | Bin 0 -> 12870 bytes assets/sendmail_template.txt | 17 + bin/markdown_to_html.py | 100 +++++ bin/markdown_to_html.r | 51 --- bin/scrape_software_versions.py | 15 +- conf/awsbatch.config | 18 - conf/base.config | 37 +- conf/igenomes.config | 451 +++++++++++++++++----- conf/test.config | 7 +- docs/images/nf-core-atacseq_logo.png | Bin 0 -> 22032 bytes docs/output.md | 4 +- docs/usage.md | 95 +++-- environment.yml | 10 +- main.nf | 278 +++++++------ nextflow.config | 56 ++- 32 files changed, 1049 insertions(+), 475 deletions(-) create mode 100644 .github/workflows/branch.yml create mode 100644 .github/workflows/ci.yml create mode 100644 .github/workflows/linting.yml delete mode 100644 .travis.yml create mode 100644 assets/nf-core-atacseq_logo.png create mode 100755 bin/markdown_to_html.py delete mode 100755 bin/markdown_to_html.r delete mode 100644 conf/awsbatch.config create mode 100644 docs/images/nf-core-atacseq_logo.png diff --git a/.github/CONTRIBUTING.md b/.github/CONTRIBUTING.md index 6376a3c9..0d1890fa 100644 --- a/.github/CONTRIBUTING.md +++ b/.github/CONTRIBUTING.md @@ -1,47 +1,57 @@ # nf-core/atacseq: Contributing Guidelines -Hi there! Many thanks for taking an interest in improving nf-core/atacseq. +Hi there! +Many thanks for taking an interest in improving nf-core/atacseq. -We try to manage the required tasks for nf-core/atacseq using GitHub issues, you probably came to this page when creating one. Please use the pre-filled template to save time. - -However, don't be put off by this template - other more general issues and suggestions are welcome! Contributions to the code are even more welcome ;) - -> If you need help using or modifying nf-core/atacseq then the best place to ask is on the pipeline channel on [Slack](https://nf-core-invite.herokuapp.com/). +We try to manage the required tasks for nf-core/atacseq using GitHub issues, you probably came to this page when creating one. +Please use the pre-filled template to save time. +However, don't be put off by this template - other more general issues and suggestions are welcome! +Contributions to the code are even more welcome ;) +> If you need help using or modifying nf-core/atacseq then the best place to ask is on the nf-core Slack [#atacseq](https://nfcore.slack.com/channels/atacseq) channel ([join our Slack here](https://nf-co.re/join/slack)). ## Contribution workflow -If you'd like to write some code for nf-core/atacseq, the standard workflow -is as follows: -1. Check that there isn't already an issue about your idea in the - [nf-core/atacseq issues](https://github.com/nf-core/atacseq/issues) to avoid - duplicating work. +If you'd like to write some code for nf-core/atacseq, the standard workflow is as follows: + +1. Check that there isn't already an issue about your idea in the [nf-core/atacseq issues](https://github.com/nf-core/atacseq/issues) to avoid duplicating work * If there isn't one already, please create one so that others know you're working on this -2. Fork the [nf-core/atacseq repository](https://github.com/nf-core/atacseq) to your GitHub account +2. [Fork](https://help.github.com/en/github/getting-started-with-github/fork-a-repo) the [nf-core/atacseq repository](https://github.com/nf-core/atacseq) to your GitHub account 3. Make the necessary changes / additions within your forked repository -4. Submit a Pull Request against the `dev` branch and wait for the code to be reviewed and merged. - -If you're not used to this workflow with git, you can start with some [basic docs from GitHub](https://help.github.com/articles/fork-a-repo/) or even their [excellent interactive tutorial](https://try.github.io/). +4. Submit a Pull Request against the `dev` branch and wait for the code to be reviewed and merged +If you're not used to this workflow with git, you can start with some [docs from GitHub](https://help.github.com/en/github/collaborating-with-issues-and-pull-requests) or even their [excellent `git` resources](https://try.github.io/). ## Tests -When you create a pull request with changes, [Travis CI](https://travis-ci.org/) will run automatic tests. + +When you create a pull request with changes, [GitHub Actions](https://github.com/features/actions) will run automatic tests. Typically, pull-requests are only fully reviewed when these tests are passing, though of course we can help out before then. There are typically two types of tests that run: ### Lint Tests -The nf-core has a [set of guidelines](http://nf-co.re/guidelines) which all pipelines must adhere to. + +`nf-core` has a [set of guidelines](https://nf-co.re/developers/guidelines) which all pipelines must adhere to. To enforce these and ensure that all pipelines stay in sync, we have developed a helper tool which runs checks on the pipeline code. This is in the [nf-core/tools repository](https://github.com/nf-core/tools) and once installed can be run locally with the `nf-core lint ` command. If any failures or warnings are encountered, please follow the listed URL for more documentation. ### Pipeline Tests -Each nf-core pipeline should be set up with a minimal set of test-data. -Travis CI then runs the pipeline on this data to ensure that it exists successfully. + +Each `nf-core` pipeline should be set up with a minimal set of test-data. +`GitHub Actions` then runs the pipeline on this data to ensure that it exits successfully. If there are any failures then the automated tests fail. -These tests are run both with the latest available version of Nextflow and also the minimum required version that is stated in the pipeline code. +These tests are run both with the latest available version of `Nextflow` and also the minimum required version that is stated in the pipeline code. + +## Patch + +: warning: Only in the unlikely and regretful event of a release happening with a bug. + +* On your own fork, make a new branch `patch` based on `upstream/master`. +* Fix the bug, and bump version (X.Y.Z+1). +* A PR should be made on `master` from patch to directly this particular bug. ## Getting help -For further information/help, please consult the [nf-core/atacseq documentation](https://github.com/nf-core/atacseq#documentation) and don't hesitate to get in touch on the pipeline channel on [Slack](https://nf-core-invite.herokuapp.com/). + +For further information/help, please consult the [nf-core/atacseq documentation](https://nf-co.re/nf-core/atacseq/docs) and don't hesitate to get in touch on the nf-core Slack [#atacseq](https://nfcore.slack.com/channels/atacseq) channel ([join our Slack here](https://nf-co.re/join/slack)). diff --git a/.github/ISSUE_TEMPLATE/bug_report.md b/.github/ISSUE_TEMPLATE/bug_report.md index b287bce7..010263cf 100644 --- a/.github/ISSUE_TEMPLATE/bug_report.md +++ b/.github/ISSUE_TEMPLATE/bug_report.md @@ -1,31 +1,42 @@ +# nf-core/atacseq bug report + Hi there! -Thanks for telling us about a problem with the pipeline. Please delete this text and anything that's not relevant from the template below: +Thanks for telling us about a problem with the pipeline. +Please delete this text and anything that's not relevant from the template below: + +## Describe the bug -#### Describe the bug A clear and concise description of what the bug is. -#### Steps to reproduce +## Steps to reproduce + Steps to reproduce the behaviour: + 1. Command line: `nextflow run ...` 2. See error: _Please provide your error message_ -#### Expected behaviour +## Expected behaviour + A clear and concise description of what you expected to happen. -#### System: - - Hardware: [e.g. HPC, Desktop, Cloud...] - - Executor: [e.g. slurm, local, awsbatch...] - - OS: [e.g. CentOS Linux, macOS, Linux Mint...] - - Version [e.g. 7, 10.13.6, 18.3...] +## System + +- Hardware: +- Executor: +- OS: +- Version + +## Nextflow Installation + +- Version: + +## Container engine -#### Nextflow Installation: - - Version: [e.g. 0.31.0] +- Engine: +- version: +- Image tag: -#### Container engine: - - Engine: [e.g. Conda, Docker or Singularity] - - version: [e.g. 1.0.0] - - Image tag: [e.g. nfcore/atacseq:1.0.0] +## Additional context -#### Additional context Add any other context about the problem here. diff --git a/.github/ISSUE_TEMPLATE/feature_request.md b/.github/ISSUE_TEMPLATE/feature_request.md index 1f025b77..222fe51a 100644 --- a/.github/ISSUE_TEMPLATE/feature_request.md +++ b/.github/ISSUE_TEMPLATE/feature_request.md @@ -1,16 +1,24 @@ +# nf-core/atacseq feature request + Hi there! -Thanks for suggesting a new feature for the pipeline! Please delete this text and anything that's not relevant from the template below: +Thanks for suggesting a new feature for the pipeline! +Please delete this text and anything that's not relevant from the template below: + +## Is your feature request related to a problem? Please describe -#### Is your feature request related to a problem? Please describe. A clear and concise description of what the problem is. + Ex. I'm always frustrated when [...] -#### Describe the solution you'd like +## Describe the solution you'd like + A clear and concise description of what you want to happen. -#### Describe alternatives you've considered +## Describe alternatives you've considered + A clear and concise description of any alternative solutions or features you've considered. -#### Additional context +## Additional context + Add any other context about the feature request here. diff --git a/.github/PULL_REQUEST_TEMPLATE.md b/.github/PULL_REQUEST_TEMPLATE.md index 2784b969..67681d0c 100644 --- a/.github/PULL_REQUEST_TEMPLATE.md +++ b/.github/PULL_REQUEST_TEMPLATE.md @@ -1,15 +1,19 @@ -Many thanks to contributing to nf-core/atacseq! +# nf-core/atacseq pull request -Please fill in the appropriate checklist below (delete whatever is not relevant). These are the most common things requested on pull requests (PRs). +Many thanks for contributing to nf-core/atacseq! + +Please fill in the appropriate checklist below (delete whatever is not relevant). +These are the most common things requested on pull requests (PRs). ## PR checklist - - [ ] This comment contains a description of changes (with reason) - - [ ] If you've fixed a bug or added code that should be tested, add tests! - - [ ] If necessary, also make a PR on the [nf-core/atacseq branch on the nf-core/test-datasets repo]( https://github.com/nf-core/test-datasets/pull/new/nf-core/atacseq) - - [ ] Ensure the test suite passes (`nextflow run . -profile test,docker`). - - [ ] Make sure your code lints (`nf-core lint .`). - - [ ] Documentation in `docs` is updated - - [ ] `CHANGELOG.md` is updated - - [ ] `README.md` is updated - -**Learn more about contributing:** https://github.com/nf-core/atacseq/tree/master/.github/CONTRIBUTING.md + +- [ ] This comment contains a description of changes (with reason) +- [ ] If you've fixed a bug or added code that should be tested, add tests! +- [ ] If necessary, also make a PR on the [nf-core/atacseq branch on the nf-core/test-datasets repo](https://github.com/nf-core/test-datasets/pull/new/nf-core/atacseq) +- [ ] Ensure the test suite passes (`nextflow run . -profile test,docker`). +- [ ] Make sure your code lints (`nf-core lint .`). +- [ ] Documentation in `docs` is updated +- [ ] `CHANGELOG.md` is updated +- [ ] `README.md` is updated + +**Learn more about contributing:** [CONTRIBUTING.md](https://github.com/nf-core/atacseq/tree/master/.github/CONTRIBUTING.md) \ No newline at end of file diff --git a/.github/markdownlint.yml b/.github/markdownlint.yml index e052a635..96b12a70 100644 --- a/.github/markdownlint.yml +++ b/.github/markdownlint.yml @@ -1,9 +1,5 @@ # Markdownlint configuration file default: true, line-length: false -no-multiple-blanks: 0 -blanks-around-headers: false -blanks-around-lists: false -header-increment: false no-duplicate-header: siblings_only: true diff --git a/.github/workflows/branch.yml b/.github/workflows/branch.yml new file mode 100644 index 00000000..9ba92076 --- /dev/null +++ b/.github/workflows/branch.yml @@ -0,0 +1,16 @@ +name: nf-core branch protection +# This workflow is triggered on PRs to master branch on the repository +# It fails when someone tries to make a PR against the nf-core `master` branch instead of `dev` +on: + pull_request: + branches: + - master + +jobs: + test: + runs-on: ubuntu-18.04 + steps: + # PRs are only ok if coming from an nf-core `dev` branch or a fork `patch` branch + - name: Check PRs + run: | + { [[ $(git remote get-url origin) == *nf-core/atacseq ]] && [[ ${GITHUB_HEAD_REF} = "dev" ]]; } || [[ ${GITHUB_HEAD_REF} == "patch" ]] diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml new file mode 100644 index 00000000..4b730381 --- /dev/null +++ b/.github/workflows/ci.yml @@ -0,0 +1,30 @@ +name: nf-core CI +# This workflow is triggered on pushes and PRs to the repository. +# It runs the pipeline with the minimal test dataset to check that it completes without any syntax errors +on: [push, pull_request] + +jobs: + test: + env: + NXF_VER: ${{ matrix.nxf_ver }} + NXF_ANSI_LOG: false + runs-on: ubuntu-latest + strategy: + matrix: + # Nextflow versions: check pipeline minimum and current latest + nxf_ver: ['19.10.0', ''] + steps: + - uses: actions/checkout@v2 + - name: Install Nextflow + run: | + wget -qO- get.nextflow.io | bash + sudo mv nextflow /usr/local/bin/ + - name: Pull docker image + run: | + docker pull nfcore/atacseq:dev + docker tag nfcore/atacseq:dev nfcore/atacseq:dev + - name: Run pipeline with test data + run: | + # TODO nf-core: You can customise CI pipeline run tests as required + # (eg. adding multiple test runs with different parameters) + nextflow run ${GITHUB_WORKSPACE} -profile test,docker diff --git a/.github/workflows/linting.yml b/.github/workflows/linting.yml new file mode 100644 index 00000000..1e0827a8 --- /dev/null +++ b/.github/workflows/linting.yml @@ -0,0 +1,50 @@ +name: nf-core linting +# This workflow is triggered on pushes and PRs to the repository. +# It runs the `nf-core lint` and markdown lint tests to ensure that the code meets the nf-core guidelines +on: + push: + pull_request: + release: + types: [published] + +jobs: + Markdown: + runs-on: ubuntu-latest + steps: + - uses: actions/checkout@v2 + - uses: actions/setup-node@v1 + with: + node-version: '10' + - name: Install markdownlint + run: npm install -g markdownlint-cli + - name: Run Markdownlint + run: markdownlint ${GITHUB_WORKSPACE} -c ${GITHUB_WORKSPACE}/.github/markdownlint.yml + YAML: + runs-on: ubuntu-latest + steps: + - uses: actions/checkout@v1 + - uses: actions/setup-node@v1 + with: + node-version: '10' + - name: Install yaml-lint + run: npm install -g yaml-lint + - name: Run yaml-lint + run: yamllint $(find ${GITHUB_WORKSPACE} -type f -name "*.yml") + nf-core: + runs-on: ubuntu-latest + steps: + - uses: actions/checkout@v2 + - name: Install Nextflow + run: | + wget -qO- get.nextflow.io | bash + sudo mv nextflow /usr/local/bin/ + - uses: actions/setup-python@v1 + with: + python-version: '3.6' + architecture: 'x64' + - name: Install dependencies + run: | + python -m pip install --upgrade pip + pip install nf-core + - name: Run nf-core lint + run: nf-core lint ${GITHUB_WORKSPACE} diff --git a/.gitignore b/.gitignore index 5b54e3e6..6354f370 100644 --- a/.gitignore +++ b/.gitignore @@ -3,5 +3,6 @@ work/ data/ results/ .DS_Store -tests/test_data +tests/ +testing/ *.pyc diff --git a/.travis.yml b/.travis.yml deleted file mode 100644 index b00e5355..00000000 --- a/.travis.yml +++ /dev/null @@ -1,42 +0,0 @@ -sudo: required -language: python -jdk: openjdk8 -services: docker -python: '3.6' -cache: pip -matrix: - fast_finish: true - -before_install: - # PRs to master are only ok if coming from dev branch - - '[ $TRAVIS_PULL_REQUEST = "false" ] || [ $TRAVIS_BRANCH != "master" ] || ([ $TRAVIS_PULL_REQUEST_SLUG = $TRAVIS_REPO_SLUG ] && [ $TRAVIS_PULL_REQUEST_BRANCH = "dev" ])' - # Pull the docker image first so the test doesn't wait for this - - docker pull nfcore/atacseq:dev - # Fake the tag locally so that the pipeline runs properly - # Looks weird when this is :dev to :dev, but makes sense when testing code for a release (:dev to :1.0.1) - - docker tag nfcore/atacseq:dev nfcore/atacseq:dev - -install: - # Install Nextflow - - mkdir /tmp/nextflow && cd /tmp/nextflow - - wget -qO- get.nextflow.io | bash - - sudo ln -s /tmp/nextflow/nextflow /usr/local/bin/nextflow - # Install nf-core/tools - - pip install --upgrade pip - - pip install nf-core - # Reset - - mkdir ${TRAVIS_BUILD_DIR}/tests && cd ${TRAVIS_BUILD_DIR}/tests - # Install markdownlint-cli - - sudo apt-get install npm && npm install -g markdownlint-cli - -env: - - NXF_VER='0.32.0' # Specify a minimum NF version that should be tested and work - - NXF_VER='' # Plus: get the latest NF version and check that it works - -script: - # Lint the pipeline code - - nf-core lint ${TRAVIS_BUILD_DIR} - # Lint the documentation - - markdownlint ${TRAVIS_BUILD_DIR} -c ${TRAVIS_BUILD_DIR}/.github/markdownlint.yml - # Run the pipeline with the test profile - - nextflow run ${TRAVIS_BUILD_DIR} -profile test,docker diff --git a/CHANGELOG.md b/CHANGELOG.md index bf57ded8..cc3d8592 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -1,4 +1,16 @@ # nf-core/atacseq: Changelog -## v1.0.0 - [date] +The format is based on [Keep a Changelog](http://keepachangelog.com/en/1.0.0/) +and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0.html). + +## v1.1.1dev - [date] + Initial release of nf-core/atacseq, created with the [nf-core](http://nf-co.re/) template. + +### `Added` + +### `Fixed` + +### `Dependencies` + +### `Deprecated` diff --git a/CODE_OF_CONDUCT.md b/CODE_OF_CONDUCT.md index 09226d0d..cf930c8a 100644 --- a/CODE_OF_CONDUCT.md +++ b/CODE_OF_CONDUCT.md @@ -34,7 +34,7 @@ This Code of Conduct applies both within project spaces and in public spaces whe ## Enforcement -Instances of abusive, harassing, or otherwise unacceptable behavior may be reported by contacting the project team on [Slack](https://nf-core-invite.herokuapp.com/). The project team will review and investigate all complaints, and will respond in a way that it deems appropriate to the circumstances. The project team is obligated to maintain confidentiality with regard to the reporter of an incident. Further details of specific enforcement policies may be posted separately. +Instances of abusive, harassing, or otherwise unacceptable behavior may be reported by contacting the project team on [Slack](https://nf-co.re/join/slack). The project team will review and investigate all complaints, and will respond in a way that it deems appropriate to the circumstances. The project team is obligated to maintain confidentiality with regard to the reporter of an incident. Further details of specific enforcement policies may be posted separately. Project maintainers who do not follow or enforce the Code of Conduct in good faith may face temporary or permanent repercussions as determined by other members of the project's leadership. diff --git a/Dockerfile b/Dockerfile index 194b4b52..e0478eae 100644 --- a/Dockerfile +++ b/Dockerfile @@ -1,7 +1,13 @@ -FROM nfcore/base +FROM nfcore/base:1.9 LABEL authors="Harshil Patel" \ - description="Docker image containing all requirements for nf-core/atacseq pipeline" + description="Docker image containing all software requirements for the nf-core/atacseq pipeline" +# Install the conda environment COPY environment.yml / RUN conda env create -f /environment.yml && conda clean -a -ENV PATH /opt/conda/envs/nf-core-atacseq-1.0.0/bin:$PATH + +# Add conda installation dir to PATH (instead of doing 'conda activate') +ENV PATH /opt/conda/envs/nf-core-atacseq-1.1.1dev/bin:$PATH + +# Dump the details of the installed packages to a file for posterity +RUN conda env export --name nf-core-atacseq-1.1.1dev > nf-core-atacseq-1.1.1dev.yml diff --git a/README.md b/README.md index 28ce2325..e763c96c 100644 --- a/README.md +++ b/README.md @@ -1,18 +1,44 @@ -# nf-core/atacseq +# ![nf-core/atacseq](docs/images/nf-core-atacseq_logo.png) **ATACSeq peak-calling and differential analysis pipeline.**. -[![Build Status](https://travis-ci.com/nf-core/atacseq.svg?branch=master)](https://travis-ci.com/nf-core/atacseq) -[![Nextflow](https://img.shields.io/badge/nextflow-%E2%89%A50.32.0-brightgreen.svg)](https://www.nextflow.io/) +[![GitHub Actions CI Status](https://github.com/nf-core/atacseq/workflows/nf-core%20CI/badge.svg)](https://github.com/nf-core/atacseq/actions) +[![GitHub Actions Linting Status](https://github.com/nf-core/atacseq/workflows/nf-core%20linting/badge.svg)](https://github.com/nf-core/atacseq/actions) +[![Nextflow](https://img.shields.io/badge/nextflow-%E2%89%A519.10.0-brightgreen.svg)](https://www.nextflow.io/) [![install with bioconda](https://img.shields.io/badge/install%20with-bioconda-brightgreen.svg)](http://bioconda.github.io/) [![Docker](https://img.shields.io/docker/automated/nfcore/atacseq.svg)](https://hub.docker.com/r/nfcore/atacseq) ## Introduction + The pipeline is built using [Nextflow](https://www.nextflow.io), a workflow tool to run tasks across multiple compute infrastructures in a very portable manner. It comes with docker containers making installation trivial and results highly reproducible. +## Quick Start + +i. Install [`nextflow`](https://nf-co.re/usage/installation) + +ii. Install either [`Docker`](https://docs.docker.com/engine/installation/) or [`Singularity`](https://www.sylabs.io/guides/3.0/user-guide/) for full pipeline reproducibility (please only use [`Conda`](https://conda.io/miniconda.html) as a last resort; see [docs](https://nf-co.re/usage/configuration#basic-configuration-profiles)) + +iii. Download the pipeline and test it on a minimal dataset with a single command + +```bash +nextflow run nf-core/atacseq -profile test, +``` + +> Please check [nf-core/configs](https://github.com/nf-core/configs#documentation) to see if a custom config file to run nf-core pipelines already exists for your Institute. If so, you can simply use `-profile ` in your command. This will enable either `docker` or `singularity` and set the appropriate execution settings for your local compute environment. + +iv. Start running your own analysis! + + + +```bash +nextflow run nf-core/atacseq -profile --reads '*_R{1,2}.fastq.gz' --genome GRCh37 +``` + +See [usage docs](docs/usage.md) for all of the available options when running the pipeline. ## Documentation + The nf-core/atacseq pipeline comes with documentation about the pipeline, found in the `docs/` directory: 1. [Installation](https://nf-co.re/usage/installation) @@ -27,4 +53,25 @@ The nf-core/atacseq pipeline comes with documentation about the pipeline, found ## Credits + nf-core/atacseq was originally written by Harshil Patel. + +## Contributions and Support + +If you would like to contribute to this pipeline, please see the [contributing guidelines](.github/CONTRIBUTING.md). + +For further information or help, don't hesitate to get in touch on [Slack](https://nfcore.slack.com/channels/atacseq) (you can join with [this invite](https://nf-co.re/join/slack)). + +## Citation + + + + +You can cite the `nf-core` publication as follows: + +> **The nf-core framework for community-curated bioinformatics pipelines.** +> +> Philip Ewels, Alexander Peltzer, Sven Fillinger, Harshil Patel, Johannes Alneberg, Andreas Wilm, Maxime Ulysse Garcia, Paolo Di Tommaso & Sven Nahnsen. +> +> _Nat Biotechnol._ 2020 Feb 13. doi: [10.1038/s41587-020-0439-x](https://dx.doi.org/10.1038/s41587-020-0439-x). +> ReadCube: [Full Access Link](https://rdcu.be/b1GjZ) diff --git a/assets/email_template.html b/assets/email_template.html index 1c4f6946..098cd4bd 100644 --- a/assets/email_template.html +++ b/assets/email_template.html @@ -11,6 +11,8 @@

+ +

nf-core/atacseq v${version}

Run Name: $runName

diff --git a/assets/email_template.txt b/assets/email_template.txt index 38059dd2..592f0991 100644 --- a/assets/email_template.txt +++ b/assets/email_template.txt @@ -1,6 +1,12 @@ -======================================== - nf-core/atacseq v${version} -======================================== +---------------------------------------------------- + ,--./,-. + ___ __ __ __ ___ /,-._.--~\\ + |\\ | |__ __ / ` / \\ |__) |__ } { + | \\| | \\__, \\__/ | \\ |___ \\`-._,-`-, + `._,._,' + nf-core/atacseq v${version} +---------------------------------------------------- + Run Name: $runName <% if (success){ diff --git a/assets/multiqc_config.yaml b/assets/multiqc_config.yaml index 8e6246d7..c00bd44c 100644 --- a/assets/multiqc_config.yaml +++ b/assets/multiqc_config.yaml @@ -3,7 +3,9 @@ report_comment: > analysis pipeline. 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+ toString(). + tokenize( '\n' )*. + toList()*. + collate( 76 )*. + collect { it.join() }. + flatten(). + join( '\n' ) %> + <% if (mqcFile){ def mqcFileObj = new File("$mqcFile") diff --git a/bin/markdown_to_html.py b/bin/markdown_to_html.py new file mode 100755 index 00000000..57cc4263 --- /dev/null +++ b/bin/markdown_to_html.py @@ -0,0 +1,100 @@ +#!/usr/bin/env python +from __future__ import print_function +import argparse +import markdown +import os +import sys + +def convert_markdown(in_fn): + input_md = open(in_fn, mode="r", encoding="utf-8").read() + html = markdown.markdown( + "[TOC]\n" + input_md, + extensions = [ + 'pymdownx.extra', + 'pymdownx.b64', + 'pymdownx.highlight', + 'pymdownx.emoji', + 'pymdownx.tilde', + 'toc' + ], + extension_configs = { + 'pymdownx.b64': { + 'base_path': os.path.dirname(in_fn) + }, + 'pymdownx.highlight': { + 'noclasses': True + }, + 'toc': { + 'title': 'Table of Contents' + } + } + ) + return html + +def wrap_html(contents): + header = """ + + + + + +
+ """ + footer = """ +
+ + + """ + return header + contents + footer + + +def parse_args(args=None): + parser = argparse.ArgumentParser() + parser.add_argument('mdfile', type=argparse.FileType('r'), nargs='?', + help='File to convert. Defaults to stdin.') + parser.add_argument('-o', '--out', type=argparse.FileType('w'), + default=sys.stdout, + help='Output file name. Defaults to stdout.') + return parser.parse_args(args) + +def main(args=None): + args = parse_args(args) + converted_md = convert_markdown(args.mdfile.name) + html = wrap_html(converted_md) + args.out.write(html) + +if __name__ == '__main__': + sys.exit(main()) diff --git a/bin/markdown_to_html.r b/bin/markdown_to_html.r deleted file mode 100755 index abe13350..00000000 --- a/bin/markdown_to_html.r +++ /dev/null @@ -1,51 +0,0 @@ -#!/usr/bin/env Rscript - -# Command line argument processing -args = commandArgs(trailingOnly=TRUE) -if (length(args) < 2) { - stop("Usage: markdown_to_html.r ", call.=FALSE) -} -markdown_fn <- args[1] -output_fn <- args[2] - -# Load / install packages -if (!require("markdown")) { - install.packages("markdown", dependencies=TRUE, repos='http://cloud.r-project.org/') - library("markdown") -} - -base_css_fn <- getOption("markdown.HTML.stylesheet") -base_css <- readChar(base_css_fn, file.info(base_css_fn)$size) -custom_css <- paste(base_css, " -body { - padding: 3em; - margin-right: 350px; - max-width: 100%; -} -#toc { - position: fixed; - right: 20px; - width: 300px; - padding-top: 20px; - overflow: scroll; - height: calc(100% - 3em - 20px); -} -#toc_header { - font-size: 1.8em; - font-weight: bold; -} -#toc > ul { - padding-left: 0; - list-style-type: none; -} -#toc > ul ul { padding-left: 20px; } -#toc > ul > li > a { display: none; } -img { max-width: 800px; } -") - -markdownToHTML( - file = markdown_fn, - output = output_fn, - stylesheet = custom_css, - options = c('toc', 'base64_images', 'highlight_code') -) diff --git a/bin/scrape_software_versions.py b/bin/scrape_software_versions.py index d66a7915..8d2ff376 100755 --- a/bin/scrape_software_versions.py +++ b/bin/scrape_software_versions.py @@ -18,14 +18,17 @@ # Search each file using its regex for k, v in regexes.items(): - with open(v[0]) as x: - versions = x.read() - match = re.search(v[1], versions) - if match: - results[k] = "v{}".format(match.group(1)) + try: + with open(v[0]) as x: + versions = x.read() + match = re.search(v[1], versions) + if match: + results[k] = "v{}".format(match.group(1)) + except IOError: + results[k] = False # Remove software set to false in results -for k in results: +for k in list(results): if not results[k]: del(results[k]) diff --git a/conf/awsbatch.config b/conf/awsbatch.config deleted file mode 100644 index 14af5866..00000000 --- a/conf/awsbatch.config +++ /dev/null @@ -1,18 +0,0 @@ -/* - * ------------------------------------------------- - * Nextflow config file for running on AWS batch - * ------------------------------------------------- - * Base config needed for running with -profile awsbatch - */ -params { - config_profile_name = 'AWSBATCH' - config_profile_description = 'AWSBATCH Cloud Profile' - config_profile_contact = 'Alexander Peltzer (@apeltzer)' - config_profile_url = 'https://aws.amazon.com/de/batch/' -} - -aws.region = params.awsregion -process.executor = 'awsbatch' -process.queue = params.awsqueue -executor.awscli = '/home/ec2-user/miniconda/bin/aws' -params.tracedir = './' diff --git a/conf/base.config b/conf/base.config index db12e92d..c9cf6f4f 100644 --- a/conf/base.config +++ b/conf/base.config @@ -13,22 +13,39 @@ process { // TODO nf-core: Check the defaults for all processes cpus = { check_max( 1 * task.attempt, 'cpus' ) } - memory = { check_max( 8.GB * task.attempt, 'memory' ) } - time = { check_max( 2.h * task.attempt, 'time' ) } + memory = { check_max( 7.GB * task.attempt, 'memory' ) } + time = { check_max( 4.h * task.attempt, 'time' ) } errorStrategy = { task.exitStatus in [143,137,104,134,139] ? 'retry' : 'finish' } maxRetries = 1 maxErrors = '-1' // Process-specific resource requirements + // NOTE - Only one of the labels below are used in the fastqc process in the main script. + // If possible, it would be nice to keep the same label naming convention when + // adding in your processes. // TODO nf-core: Customise requirements for specific processes. // See https://www.nextflow.io/docs/latest/config.html#config-process-selectors -} - -params { - // Defaults only, expecting to be overwritten - max_memory = 128.GB - max_cpus = 16 - max_time = 240.h - igenomes_base = 's3://ngi-igenomes/igenomes/' + withLabel:process_low { + cpus = { check_max( 2 * task.attempt, 'cpus' ) } + memory = { check_max( 14.GB * task.attempt, 'memory' ) } + time = { check_max( 6.h * task.attempt, 'time' ) } + } + withLabel:process_medium { + cpus = { check_max( 6 * task.attempt, 'cpus' ) } + memory = { check_max( 42.GB * task.attempt, 'memory' ) } + time = { check_max( 8.h * task.attempt, 'time' ) } + } + withLabel:process_high { + cpus = { check_max( 12 * task.attempt, 'cpus' ) } + memory = { check_max( 84.GB * task.attempt, 'memory' ) } + time = { check_max( 10.h * task.attempt, 'time' ) } + } + withLabel:process_long { + time = { check_max( 20.h * task.attempt, 'time' ) } + } + withName:get_software_versions { + cache = false + } + } diff --git a/conf/igenomes.config b/conf/igenomes.config index d19e61f4..2de92422 100644 --- a/conf/igenomes.config +++ b/conf/igenomes.config @@ -9,139 +9,412 @@ params { // illumina iGenomes reference file paths - // TODO nf-core: Add new reference types and strip out those that are not needed genomes { 'GRCh37' { - bed12 = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Annotation/Genes/genes.bed" - fasta = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/WholeGenomeFasta/genome.fa" - gtf = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Annotation/Genes/genes.gtf" - star = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/STARIndex/" + fasta = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Annotation/README.txt" + mito_name = "MT" + macs_gsize = "2.7e9" + blacklist = "${baseDir}/assets/blacklists/GRCh37-blacklist.bed" + } + 'GRCh38' { + fasta = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Annotation/Genes/genes.bed" + mito_name = "chrM" + macs_gsize = "2.7e9" + blacklist = "${baseDir}/assets/blacklists/hg38-blacklist.bed" } 'GRCm38' { - bed12 = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Annotation/Genes/genes.bed" - fasta = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/WholeGenomeFasta/genome.fa" - gtf = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Annotation/Genes/genes.gtf" - star = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/STARIndex/" + fasta = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Annotation/README.txt" + mito_name = "MT" + macs_gsize = "1.87e9" + blacklist = "${baseDir}/assets/blacklists/GRCm38-blacklist.bed" } 'TAIR10' { - bed12 = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Annotation/Genes/genes.bed" - fasta = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/WholeGenomeFasta/genome.fa" - gtf = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Annotation/Genes/genes.gtf" - star = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/STARIndex/" + fasta = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Annotation/README.txt" + mito_name = "Mt" } 'EB2' { - bed12 = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Annotation/Genes/genes.bed" - fasta = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/WholeGenomeFasta/genome.fa" - gtf = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Annotation/Genes/genes.gtf" - star = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/STARIndex/" + fasta = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Annotation/README.txt" } 'UMD3.1' { - bed12 = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Annotation/Genes/genes.bed" - fasta = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/WholeGenomeFasta/genome.fa" - gtf = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Annotation/Genes/genes.gtf" - star = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/STARIndex/" + fasta = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Annotation/README.txt" + mito_name = "MT" } 'WBcel235' { - bed12 = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Annotation/Genes/genes.bed" - fasta = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/WholeGenomeFasta/genome.fa" - gtf = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Annotation/Genes/genes.gtf" - star = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/STARIndex/" + fasta = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Annotation/Genes/genes.bed" + mito_name = "MtDNA" + macs_gsize = "9e7" } 'CanFam3.1' { - bed12 = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Annotation/Genes/genes.bed" - fasta = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/WholeGenomeFasta/genome.fa" - gtf = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Annotation/Genes/genes.gtf" - star = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/STARIndex/" + fasta = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Annotation/README.txt" + mito_name = "MT" } 'GRCz10' { - bed12 = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Annotation/Genes/genes.bed" - fasta = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/WholeGenomeFasta/genome.fa" - gtf = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Annotation/Genes/genes.gtf" - star = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/STARIndex/" + fasta = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Annotation/Genes/genes.bed" + mito_name = "MT" } 'BDGP6' { - bed12 = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Annotation/Genes/genes.bed" - fasta = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/WholeGenomeFasta/genome.fa" - gtf = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Annotation/Genes/genes.gtf" - star = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/STARIndex/" + fasta = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Annotation/Genes/genes.bed" + mito_name = "M" + macs_gsize = "1.2e8" } 'EquCab2' { - bed12 = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Annotation/Genes/genes.bed" - fasta = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/WholeGenomeFasta/genome.fa" - gtf = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Annotation/Genes/genes.gtf" - star = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/STARIndex/" + fasta = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Annotation/README.txt" + mito_name = "MT" } 'EB1' { - bed12 = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Annotation/Genes/genes.bed" - fasta = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/WholeGenomeFasta/genome.fa" - gtf = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Annotation/Genes/genes.gtf" - star = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/STARIndex/" + fasta = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Annotation/README.txt" } 'Galgal4' { - bed12 = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Annotation/Genes/genes.bed" - fasta = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/WholeGenomeFasta/genome.fa" - gtf = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Annotation/Genes/genes.gtf" - star = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/STARIndex/" + fasta = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Annotation/Genes/genes.bed" + mito_name = "MT" } 'Gm01' { - bed12 = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Annotation/Genes/genes.bed" - fasta = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/WholeGenomeFasta/genome.fa" - gtf = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Annotation/Genes/genes.gtf" - star = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/STARIndex/" + fasta = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Annotation/README.txt" } 'Mmul_1' { - bed12 = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Annotation/Genes/genes.bed" - fasta = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/WholeGenomeFasta/genome.fa" - gtf = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Annotation/Genes/genes.gtf" - star = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/STARIndex/" + fasta = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Annotation/README.txt" + mito_name = "MT" } 'IRGSP-1.0' { - bed12 = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Annotation/Genes/genes.bed" - fasta = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/WholeGenomeFasta/genome.fa" - gtf = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Annotation/Genes/genes.gtf" - star = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/STARIndex/" + fasta = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Annotation/Genes/genes.bed" + mito_name = "Mt" } 'CHIMP2.1.4' { - bed12 = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Annotation/Genes/genes.bed" - fasta = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/WholeGenomeFasta/genome.fa" - gtf = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Annotation/Genes/genes.gtf" - star = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/STARIndex/" + fasta = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Annotation/README.txt" + mito_name = "MT" } 'Rnor_6.0' { - bed12 = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Annotation/Genes/genes.bed" - fasta = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/WholeGenomeFasta/genome.fa" - gtf = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Annotation/Genes/genes.gtf" - star = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/STARIndex/" + fasta = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Annotation/Genes/genes.bed" + mito_name = "MT" } 'R64-1-1' { - bed12 = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Annotation/Genes/genes.bed" - fasta = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/WholeGenomeFasta/genome.fa" - gtf = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Annotation/Genes/genes.gtf" - star = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/STARIndex/" + fasta = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Annotation/Genes/genes.bed" + mito_name = "MT" + macs_gsize = "1.2e7" } 'EF2' { - bed12 = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Annotation/Genes/genes.bed" - fasta = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/WholeGenomeFasta/genome.fa" - gtf = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Annotation/Genes/genes.gtf" - star = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/STARIndex/" + fasta = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Annotation/README.txt" + mito_name = "MT" + macs_gsize = "1.21e7" } 'Sbi1' { - bed12 = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Annotation/Genes/genes.bed" - fasta = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/WholeGenomeFasta/genome.fa" - gtf = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Annotation/Genes/genes.gtf" - star = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/STARIndex/" + fasta = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Annotation/README.txt" } 'Sscrofa10.2' { - bed12 = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Annotation/Genes/genes.bed" - fasta = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/WholeGenomeFasta/genome.fa" - gtf = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Annotation/Genes/genes.gtf" - star = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/STARIndex/" + fasta = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Annotation/README.txt" + mito_name = "MT" } 'AGPv3' { - bed12 = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Annotation/Genes/genes.bed" - fasta = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/WholeGenomeFasta/genome.fa" - gtf = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Annotation/Genes/genes.gtf" - star = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/STARIndex/" + fasta = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Annotation/Genes/genes.bed" + mito_name = "Mt" + } + 'hg38' { + fasta = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Annotation/Genes/genes.bed" + mito_name = "chrM" + macs_gsize = "2.7e9" + blacklist = "${baseDir}/assets/blacklists/hg38-blacklist.bed" + } + 'hg19' { + fasta = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Annotation/README.txt" + mito_name = "chrM" + macs_gsize = "2.7e9" + blacklist = "${baseDir}/assets/blacklists/hg19-blacklist.bed" + } + 'mm10' { + fasta = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Annotation/README.txt" + mito_name = "chrM" + macs_gsize = "1.87e9" + blacklist = "${baseDir}/assets/blacklists/mm10-blacklist.bed" + } + 'bosTau8' { + fasta = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Annotation/Genes/genes.bed" + mito_name = "chrM" + } + 'ce10' { + fasta = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Annotation/README.txt" + mito_name = "chrM" + macs_gsize = "9e7" + } + 'canFam3' { + fasta = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Annotation/README.txt" + mito_name = "chrM" + } + 'danRer10' { + fasta = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Annotation/Genes/genes.bed" + mito_name = "chrM" + } + 'dm6' { + fasta = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Annotation/Genes/genes.bed" + mito_name = "chrM" + macs_gsize = "1.2e8" + } + 'equCab2' { + fasta = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Annotation/README.txt" + mito_name = "chrM" + } + 'galGal4' { + fasta = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Annotation/README.txt" + mito_name = "chrM" + } + 'panTro4' { + fasta = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Annotation/README.txt" + mito_name = "chrM" + } + 'rn6' { + fasta = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Annotation/Genes/genes.bed" + mito_name = "chrM" + } + 'sacCer3' { + fasta = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/BismarkIndex/" + readme = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Annotation/README.txt" + mito_name = "chrM" + macs_gsize = "1.2e7" + } + 'susScr3' { + fasta = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Annotation/Genes/genes.bed" + readme = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Annotation/README.txt" + mito_name = "chrM" } } } diff --git a/conf/test.config b/conf/test.config index c77002df..afce8701 100644 --- a/conf/test.config +++ b/conf/test.config @@ -4,20 +4,21 @@ * ------------------------------------------------- * Defines bundled input files and everything required * to run a fast and simple test. Use as follows: - * nextflow run nf-core/atacseq -profile test + * nextflow run nf-core/atacseq -profile test, */ params { config_profile_name = 'Test profile' config_profile_description = 'Minimal test dataset to check pipeline function' - // Limit resources so that this can run on Travis + // Limit resources so that this can run on GitHub Actions max_cpus = 2 max_memory = 6.GB max_time = 48.h + // Input data // TODO nf-core: Specify the paths to your test data on nf-core/test-datasets // TODO nf-core: Give any required params for the test so that command line flags are not needed - singleEnd = false + single_end = false readPaths = [ ['Testdata', ['https://github.com/nf-core/test-datasets/raw/exoseq/testdata/Testdata_R1.tiny.fastq.gz', 'https://github.com/nf-core/test-datasets/raw/exoseq/testdata/Testdata_R2.tiny.fastq.gz']], ['SRR389222', ['https://github.com/nf-core/test-datasets/raw/methylseq/testdata/SRR389222_sub1.fastq.gz', 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Most of the plots a ## Pipeline overview + The pipeline is built using [Nextflow](https://www.nextflow.io/) and processes data using the following steps: @@ -12,6 +13,7 @@ and processes data using the following steps: * [MultiQC](#multiqc) - aggregate report, describing results of the whole pipeline ## FastQC + [FastQC](http://www.bioinformatics.babraham.ac.uk/projects/fastqc/) gives general quality metrics about your reads. It provides information about the quality score distribution across your reads, the per base sequence content (%T/A/G/C). You get information about adapter contamination and other overrepresented sequences. For further reading and documentation see the [FastQC help](http://www.bioinformatics.babraham.ac.uk/projects/fastqc/Help/). @@ -25,8 +27,8 @@ For further reading and documentation see the [FastQC help](http://www.bioinform * `zips/sample_fastqc.zip` * zip file containing the FastQC report, tab-delimited data file and plot images - ## MultiQC + [MultiQC](http://multiqc.info) is a visualisation tool that generates a single HTML report summarising all samples in your project. Most of the pipeline QC results are visualised in the report and further statistics are available in within the report data directory. The pipeline has special steps which allow the software versions used to be reported in the MultiQC output for future traceability. diff --git a/docs/usage.md b/docs/usage.md index 7f9b905b..9257b8e8 100644 --- a/docs/usage.md +++ b/docs/usage.md @@ -2,8 +2,6 @@ ## Table of contents - - * [Table of contents](#table-of-contents) * [Introduction](#introduction) * [Running the pipeline](#running-the-pipeline) @@ -12,20 +10,23 @@ * [Main arguments](#main-arguments) * [`-profile`](#-profile) * [`--reads`](#--reads) - * [`--singleEnd`](#--singleend) + * [`--single_end`](#--single_end) * [Reference genomes](#reference-genomes) * [`--genome` (using iGenomes)](#--genome-using-igenomes) * [`--fasta`](#--fasta) - * [`--igenomesIgnore`](#--igenomesignore) + * [`--igenomes_ignore`](#--igenomes_ignore) * [Job resources](#job-resources) * [Automatic resubmission](#automatic-resubmission) * [Custom resource requests](#custom-resource-requests) * [AWS Batch specific parameters](#aws-batch-specific-parameters) * [`--awsqueue`](#--awsqueue) * [`--awsregion`](#--awsregion) + * [`--awscli`](#--awscli) * [Other command line parameters](#other-command-line-parameters) * [`--outdir`](#--outdir) * [`--email`](#--email) + * [`--email_on_fail`](#--email_on_fail) + * [`--max_multiqc_email_size`](#--max_multiqc_email_size) * [`-name`](#-name) * [`-resume`](#-resume) * [`-c`](#-c) @@ -37,10 +38,9 @@ * [`--plaintext_email`](#--plaintext_email) * [`--monochrome_logs`](#--monochrome_logs) * [`--multiqc_config`](#--multiqc_config) - - ## Introduction + Nextflow handles job submissions on SLURM or other environments, and supervises running the jobs. Thus the Nextflow process must run until the pipeline is finished. We recommend that you put the process running in the background through `screen` / `tmux` or similar tool. Alternatively you can run nextflow within a cluster job submitted your job scheduler. It is recommended to limit the Nextflow Java virtual machines memory. We recommend adding the following line to your environment (typically in `~/.bashrc` or `~./bash_profile`): @@ -52,6 +52,7 @@ NXF_OPTS='-Xms1g -Xmx4g' ## Running the pipeline + The typical command for running the pipeline is as follows: ```bash @@ -70,6 +71,7 @@ results # Finished results (configurable, see below) ``` ### Updating the pipeline + When you run the above command, Nextflow automatically pulls the pipeline code from GitHub and stores it as a cached version. When running the pipeline after this, it will always use the cached version if available - even if the pipeline has been updated since. To make sure that you're running the latest version of the pipeline, make sure that you regularly update the cached version of the pipeline: ```bash @@ -77,31 +79,40 @@ nextflow pull nf-core/atacseq ``` ### Reproducibility + It's a good idea to specify a pipeline version when running the pipeline on your data. This ensures that a specific version of the pipeline code and software are used when you run your pipeline. If you keep using the same tag, you'll be running the same version of the pipeline, even if there have been changes to the code since. First, go to the [nf-core/atacseq releases page](https://github.com/nf-core/atacseq/releases) and find the latest version number - numeric only (eg. `1.3.1`). Then specify this when running the pipeline with `-r` (one hyphen) - eg. `-r 1.3.1`. This version number will be logged in reports when you run the pipeline, so that you'll know what you used when you look back in the future. - ## Main arguments ### `-profile` -Use this parameter to choose a configuration profile. Profiles can give configuration presets for different compute environments. Note that multiple profiles can be loaded, for example: `-profile docker` - the order of arguments is important! -If `-profile` is not specified at all the pipeline will be run locally and expects all software to be installed and available on the `PATH`. +Use this parameter to choose a configuration profile. Profiles can give configuration presets for different compute environments. + +Several generic profiles are bundled with the pipeline which instruct the pipeline to use software packaged using different methods (Docker, Singularity, Conda) - see below. + +> We highly recommend the use of Docker or Singularity containers for full pipeline reproducibility, however when this is not possible, Conda is also supported. + +The pipeline also dynamically loads configurations from [https://github.com/nf-core/configs](https://github.com/nf-core/configs) when it runs, making multiple config profiles for various institutional clusters available at run time. For more information and to see if your system is available in these configs please see the [nf-core/configs documentation](https://github.com/nf-core/configs#documentation). + +Note that multiple profiles can be loaded, for example: `-profile test,docker` - the order of arguments is important! +They are loaded in sequence, so later profiles can overwrite earlier profiles. + +If `-profile` is not specified, the pipeline will run locally and expect all software to be installed and available on the `PATH`. This is _not_ recommended. -* `awsbatch` - * A generic configuration profile to be used with AWS Batch. -* `conda` - * A generic configuration profile to be used with [conda](https://conda.io/docs/) - * Pulls most software from [Bioconda](https://bioconda.github.io/) * `docker` * A generic configuration profile to be used with [Docker](http://docker.com/) * Pulls software from dockerhub: [`nfcore/atacseq`](http://hub.docker.com/r/nfcore/atacseq/) * `singularity` * A generic configuration profile to be used with [Singularity](http://singularity.lbl.gov/) * Pulls software from DockerHub: [`nfcore/atacseq`](http://hub.docker.com/r/nfcore/atacseq/) +* `conda` + * Please only use Conda as a last resort i.e. when it's not possible to run the pipeline with Docker or Singularity. + * A generic configuration profile to be used with [Conda](https://conda.io/docs/) + * Pulls most software from [Bioconda](https://bioconda.github.io/) * `test` * A profile with a complete configuration for automated testing * Includes links to test data so needs no other parameters @@ -109,6 +120,7 @@ If `-profile` is not specified at all the pipeline will be run locally and expec ### `--reads` + Use this to specify the location of your input FastQ files. For example: ```bash @@ -123,21 +135,22 @@ Please note the following requirements: If left unspecified, a default pattern is used: `data/*{1,2}.fastq.gz` -### `--singleEnd` -By default, the pipeline expects paired-end data. If you have single-end data, you need to specify `--singleEnd` on the command line when you launch the pipeline. A normal glob pattern, enclosed in quotation marks, can then be used for `--reads`. For example: +### `--single_end` + +By default, the pipeline expects paired-end data. If you have single-end data, you need to specify `--single_end` on the command line when you launch the pipeline. A normal glob pattern, enclosed in quotation marks, can then be used for `--reads`. For example: ```bash ---singleEnd --reads '*.fastq' +--single_end --reads '*.fastq' ``` It is not possible to run a mixture of single-end and paired-end files in one run. - ## Reference genomes The pipeline config files come bundled with paths to the illumina iGenomes reference index files. If running with docker or AWS, the configuration is set up to use the [AWS-iGenomes](https://ewels.github.io/AWS-iGenomes/) resource. ### `--genome` (using iGenomes) + There are 31 different species supported in the iGenomes references. To run the pipeline, you must specify which to use with the `--genome` flag. You can find the keys to specify the genomes in the [iGenomes config file](../conf/igenomes.config). Common genomes that are supported are: @@ -171,33 +184,48 @@ params { ``` + ### `--fasta` + If you prefer, you can specify the full path to your reference genome when you run the pipeline: ```bash --fasta '[path to Fasta reference]' ``` -### `--igenomesIgnore` +### `--igenomes_ignore` + Do not load `igenomes.config` when running the pipeline. You may choose this option if you observe clashes between custom parameters and those supplied in `igenomes.config`. ## Job resources + ### Automatic resubmission + Each step in the pipeline has a default set of requirements for number of CPUs, memory and time. For most of the steps in the pipeline, if the job exits with an error code of `143` (exceeded requested resources) it will automatically resubmit with higher requests (2 x original, then 3 x original). If it still fails after three times then the pipeline is stopped. ### Custom resource requests + Wherever process-specific requirements are set in the pipeline, the default value can be changed by creating a custom config file. See the files hosted at [`nf-core/configs`](https://github.com/nf-core/configs/tree/master/conf) for examples. If you are likely to be running `nf-core` pipelines regularly it may be a good idea to request that your custom config file is uploaded to the `nf-core/configs` git repository. Before you do this please can you test that the config file works with your pipeline of choice using the `-c` parameter (see definition below). You can then create a pull request to the `nf-core/configs` repository with the addition of your config file, associated documentation file (see examples in [`nf-core/configs/docs`](https://github.com/nf-core/configs/tree/master/docs)), and amending [`nfcore_custom.config`](https://github.com/nf-core/configs/blob/master/nfcore_custom.config) to include your custom profile. -If you have any questions or issues please send us a message on [Slack](https://nf-core-invite.herokuapp.com/). +If you have any questions or issues please send us a message on [Slack](https://nf-co.re/join/slack). ## AWS Batch specific parameters -Running the pipeline on AWS Batch requires a couple of specific parameters to be set according to your AWS Batch configuration. Please use the `-awsbatch` profile and then specify all of the following parameters. + +Running the pipeline on AWS Batch requires a couple of specific parameters to be set according to your AWS Batch configuration. Please use [`-profile awsbatch`](https://github.com/nf-core/configs/blob/master/conf/awsbatch.config) and then specify all of the following parameters. + ### `--awsqueue` + The JobQueue that you intend to use on AWS Batch. + ### `--awsregion` -The AWS region to run your job in. Default is set to `eu-west-1` but can be adjusted to your needs. + +The AWS region in which to run your job. Default is set to `eu-west-1` but can be adjusted to your needs. + +### `--awscli` + +The [AWS CLI](https://www.nextflow.io/docs/latest/awscloud.html#aws-cli-installation) path in your custom AMI. Default: `/home/ec2-user/miniconda/bin/aws`. Please make sure to also set the `-w/--work-dir` and `--outdir` parameters to a S3 storage bucket of your choice - you'll get an error message notifying you if you didn't. @@ -206,12 +234,23 @@ Please make sure to also set the `-w/--work-dir` and `--outdir` parameters to a ### `--outdir` + The output directory where the results will be saved. ### `--email` + Set this parameter to your e-mail address to get a summary e-mail with details of the run sent to you when the workflow exits. If set in your user config file (`~/.nextflow/config`) then you don't need to specify this on the command line for every run. +### `--email_on_fail` + +This works exactly as with `--email`, except emails are only sent if the workflow is not successful. + +### `--max_multiqc_email_size` + +Threshold size for MultiQC report to be attached in notification email. If file generated by pipeline exceeds the threshold, it will not be attached (Default: 25MB). + ### `-name` + Name for the pipeline run. If not specified, Nextflow will automatically generate a random mnemonic. This is used in the MultiQC report (if not default) and in the summary HTML / e-mail (always). @@ -219,6 +258,7 @@ This is used in the MultiQC report (if not default) and in the summary HTML / e- **NB:** Single hyphen (core Nextflow option) ### `-resume` + Specify this when restarting a pipeline. Nextflow will used cached results from any pipeline steps where the inputs are the same, continuing from where it got to previously. You can also supply a run name to resume a specific run: `-resume [run-name]`. Use the `nextflow log` command to show previous run names. @@ -226,6 +266,7 @@ You can also supply a run name to resume a specific run: `-resume [run-name]`. U **NB:** Single hyphen (core Nextflow option) ### `-c` + Specify the path to a specific config file (this is a core NextFlow command). **NB:** Single hyphen (core Nextflow option) @@ -233,7 +274,8 @@ Specify the path to a specific config file (this is a core NextFlow command). Note - you can use this to override pipeline defaults. ### `--custom_config_version` -Provide git commit id for custom Institutional configs hosted at `nf-core/configs`. This was implemented for reproducibility purposes. Default is set to `master`. + +Provide git commit id for custom Institutional configs hosted at `nf-core/configs`. This was implemented for reproducibility purposes. Default: `master`. ```bash ## Download and use config file with following git commid id @@ -241,6 +283,7 @@ Provide git commit id for custom Institutional configs hosted at `nf-core/config ``` ### `--custom_config_base` + If you're running offline, nextflow will not be able to fetch the institutional config files from the internet. If you don't need them, then this is not a problem. If you do need them, you should download the files from the repo and tell nextflow where to find them with the @@ -261,22 +304,28 @@ nextflow run /path/to/pipeline/ --custom_config_base /path/to/my/configs/configs > files + singularity containers + institutional configs in one go for you, to make this process easier. ### `--max_memory` + Use to set a top-limit for the default memory requirement for each process. Should be a string in the format integer-unit. eg. `--max_memory '8.GB'` ### `--max_time` + Use to set a top-limit for the default time requirement for each process. Should be a string in the format integer-unit. eg. `--max_time '2.h'` ### `--max_cpus` + Use to set a top-limit for the default CPU requirement for each process. Should be a string in the format integer-unit. eg. `--max_cpus 1` ### `--plaintext_email` + Set to receive plain-text e-mails instead of HTML formatted. ### `--monochrome_logs` + Set to disable colourful command line output and live life in monochrome. ### `--multiqc_config` + Specify a path to a custom MultiQC configuration file. diff --git a/environment.yml b/environment.yml index 2fe4a741..e20eb85f 100644 --- a/environment.yml +++ b/environment.yml @@ -1,11 +1,15 @@ # You can use this file to create a conda environment for this pipeline: # conda env create -f environment.yml -name: nf-core-atacseq-1.0.0 +name: nf-core-atacseq-1.1.1dev channels: - conda-forge - bioconda - defaults dependencies: + - conda-forge::python=3.7.3 + - conda-forge::markdown=3.1.1 + - conda-forge::pymdown-extensions=6.0 + - conda-forge::pygments=2.5.2 # TODO nf-core: Add required software dependencies here - - fastqc=0.11.8 - - multiqc=1.7 + - bioconda::fastqc=0.11.8 + - bioconda::multiqc=1.7 diff --git a/main.nf b/main.nf index ff53c27e..0c5f289b 100644 --- a/main.nf +++ b/main.nf @@ -9,7 +9,6 @@ ---------------------------------------------------------------------------------------- */ - def helpMessage() { // TODO nf-core: Add to this help message with new command line parameters log.info nfcoreHeader() @@ -22,39 +21,41 @@ def helpMessage() { nextflow run nf-core/atacseq --reads '*_R{1,2}.fastq.gz' -profile docker Mandatory arguments: - --reads Path to input data (must be surrounded with quotes) - -profile Configuration profile to use. Can use multiple (comma separated) - Available: conda, docker, singularity, awsbatch, test and more. + --reads [file] Path to input data (must be surrounded with quotes) + -profile [str] Configuration profile to use. Can use multiple (comma separated) + Available: conda, docker, singularity, test, awsbatch, and more Options: - --genome Name of iGenomes reference - --singleEnd Specifies that the input is single end reads + --genome [str] Name of iGenomes reference + --single_end [bool] Specifies that the input is single-end reads - References If not specified in the configuration file or you wish to overwrite any of the references. - --fasta Path to Fasta reference + References If not specified in the configuration file or you wish to overwrite any of the references + --fasta [file] Path to fasta reference Other options: - --outdir The output directory where the results will be saved - --email Set this parameter to your e-mail address to get a summary e-mail with details of the run sent to you when the workflow exits - --maxMultiqcEmailFileSize Theshold size for MultiQC report to be attached in notification email. If file generated by pipeline exceeds the threshold, it will not be attached (Default: 25MB) - -name Name for the pipeline run. If not specified, Nextflow will automatically generate a random mnemonic. + --outdir [file] The output directory where the results will be saved + --email [email] Set this parameter to your e-mail address to get a summary e-mail with details of the run sent to you when the workflow exits + --email_on_fail [email] Same as --email, except only send mail if the workflow is not successful + --max_multiqc_email_size [str] Theshold size for MultiQC report to be attached in notification email. If file generated by pipeline exceeds the threshold, it will not be attached (Default: 25MB) + -name [str] Name for the pipeline run. If not specified, Nextflow will automatically generate a random mnemonic AWSBatch options: - --awsqueue The AWSBatch JobQueue that needs to be set when running on AWSBatch - --awsregion The AWS Region for your AWS Batch job to run on + --awsqueue [str] The AWSBatch JobQueue that needs to be set when running on AWSBatch + --awsregion [str] The AWS Region for your AWS Batch job to run on + --awscli [str] Path to the AWS CLI tool """.stripIndent() } -/* - * SET UP CONFIGURATION VARIABLES - */ - -// Show help emssage -if (params.help){ +// Show help message +if (params.help) { helpMessage() exit 0 } +/* + * SET UP CONFIGURATION VARIABLES + */ + // Check if genome exists in the config file if (params.genomes && params.genome && !params.genomes.containsKey(params.genome)) { exit 1, "The provided genome '${params.genome}' is not available in the iGenomes file. Currently the available genomes are ${params.genomes.keySet().join(", ")}" @@ -62,103 +63,101 @@ if (params.genomes && params.genome && !params.genomes.containsKey(params.genome // TODO nf-core: Add any reference files that are needed // Configurable reference genomes -fasta = params.genome ? params.genomes[ params.genome ].fasta ?: false : false -if ( params.fasta ){ - fasta = file(params.fasta) - if( !fasta.exists() ) exit 1, "Fasta file not found: ${params.fasta}" -} // // NOTE - THIS IS NOT USED IN THIS PIPELINE, EXAMPLE ONLY -// If you want to use the above in a process, define the following: +// If you want to use the channel below in a process, define the following: // input: -// file fasta from fasta +// file fasta from ch_fasta // - +params.fasta = params.genome ? params.genomes[ params.genome ].fasta ?: false : false +if (params.fasta) { ch_fasta = file(params.fasta, checkIfExists: true) } // Has the run name been specified by the user? // this has the bonus effect of catching both -name and --name custom_runName = params.name -if( !(workflow.runName ==~ /[a-z]+_[a-z]+/) ){ - custom_runName = workflow.runName +if (!(workflow.runName ==~ /[a-z]+_[a-z]+/)) { + custom_runName = workflow.runName } - -if( workflow.profile == 'awsbatch') { - // AWSBatch sanity checking - if (!params.awsqueue || !params.awsregion) exit 1, "Specify correct --awsqueue and --awsregion parameters on AWSBatch!" - // Check outdir paths to be S3 buckets if running on AWSBatch - // related: https://github.com/nextflow-io/nextflow/issues/813 - if (!params.outdir.startsWith('s3:')) exit 1, "Outdir not on S3 - specify S3 Bucket to run on AWSBatch!" - // Prevent trace files to be stored on S3 since S3 does not support rolling files. - if (workflow.tracedir.startsWith('s3:')) exit 1, "Specify a local tracedir or run without trace! S3 cannot be used for tracefiles." +if (workflow.profile.contains('awsbatch')) { + // AWSBatch sanity checking + if (!params.awsqueue || !params.awsregion) exit 1, "Specify correct --awsqueue and --awsregion parameters on AWSBatch!" + // Check outdir paths to be S3 buckets if running on AWSBatch + // related: https://github.com/nextflow-io/nextflow/issues/813 + if (!params.outdir.startsWith('s3:')) exit 1, "Outdir not on S3 - specify S3 Bucket to run on AWSBatch!" + // Prevent trace files to be stored on S3 since S3 does not support rolling files. + if (params.tracedir.startsWith('s3:')) exit 1, "Specify a local tracedir or run without trace! S3 cannot be used for tracefiles." } // Stage config files -ch_multiqc_config = Channel.fromPath(params.multiqc_config) -ch_output_docs = Channel.fromPath("$baseDir/docs/output.md") +ch_multiqc_config = file("$baseDir/assets/multiqc_config.yaml", checkIfExists: true) +ch_multiqc_custom_config = params.multiqc_config ? Channel.fromPath(params.multiqc_config, checkIfExists: true) : Channel.empty() +ch_output_docs = file("$baseDir/docs/output.md", checkIfExists: true) /* * Create a channel for input read files */ -if(params.readPaths){ - if(params.singleEnd){ +if (params.readPaths) { + if (params.single_end) { Channel .from(params.readPaths) - .map { row -> [ row[0], [file(row[1][0])]] } + .map { row -> [ row[0], [ file(row[1][0], checkIfExists: true) ] ] } .ifEmpty { exit 1, "params.readPaths was empty - no input files supplied" } - .into { read_files_fastqc; read_files_trimming } + .into { ch_read_files_fastqc; ch_read_files_trimming } } else { Channel .from(params.readPaths) - .map { row -> [ row[0], [file(row[1][0]), file(row[1][1])]] } + .map { row -> [ row[0], [ file(row[1][0], checkIfExists: true), file(row[1][1], checkIfExists: true) ] ] } .ifEmpty { exit 1, "params.readPaths was empty - no input files supplied" } - .into { read_files_fastqc; read_files_trimming } + .into { ch_read_files_fastqc; ch_read_files_trimming } } } else { Channel - .fromFilePairs( params.reads, size: params.singleEnd ? 1 : 2 ) - .ifEmpty { exit 1, "Cannot find any reads matching: ${params.reads}\nNB: Path needs to be enclosed in quotes!\nIf this is single-end data, please specify --singleEnd on the command line." } - .into { read_files_fastqc; read_files_trimming } + .fromFilePairs(params.reads, size: params.single_end ? 1 : 2) + .ifEmpty { exit 1, "Cannot find any reads matching: ${params.reads}\nNB: Path needs to be enclosed in quotes!\nIf this is single-end data, please specify --single_end on the command line." } + .into { ch_read_files_fastqc; ch_read_files_trimming } } - // Header log info log.info nfcoreHeader() def summary = [:] -if(workflow.revision) summary['Pipeline Release'] = workflow.revision +if (workflow.revision) summary['Pipeline Release'] = workflow.revision summary['Run Name'] = custom_runName ?: workflow.runName // TODO nf-core: Report custom parameters here summary['Reads'] = params.reads summary['Fasta Ref'] = params.fasta -summary['Data Type'] = params.singleEnd ? 'Single-End' : 'Paired-End' +summary['Data Type'] = params.single_end ? 'Single-End' : 'Paired-End' summary['Max Resources'] = "$params.max_memory memory, $params.max_cpus cpus, $params.max_time time per job" -if(workflow.containerEngine) summary['Container'] = "$workflow.containerEngine - $workflow.container" +if (workflow.containerEngine) summary['Container'] = "$workflow.containerEngine - $workflow.container" summary['Output dir'] = params.outdir summary['Launch dir'] = workflow.launchDir summary['Working dir'] = workflow.workDir summary['Script dir'] = workflow.projectDir summary['User'] = workflow.userName -if(workflow.profile == 'awsbatch'){ - summary['AWS Region'] = params.awsregion - summary['AWS Queue'] = params.awsqueue +if (workflow.profile.contains('awsbatch')) { + summary['AWS Region'] = params.awsregion + summary['AWS Queue'] = params.awsqueue + summary['AWS CLI'] = params.awscli } summary['Config Profile'] = workflow.profile -if(params.config_profile_description) summary['Config Description'] = params.config_profile_description -if(params.config_profile_contact) summary['Config Contact'] = params.config_profile_contact -if(params.config_profile_url) summary['Config URL'] = params.config_profile_url -if(params.email) { - summary['E-mail Address'] = params.email - summary['MultiQC maxsize'] = params.maxMultiqcEmailFileSize +if (params.config_profile_description) summary['Config Description'] = params.config_profile_description +if (params.config_profile_contact) summary['Config Contact'] = params.config_profile_contact +if (params.config_profile_url) summary['Config URL'] = params.config_profile_url +if (params.email || params.email_on_fail) { + summary['E-mail Address'] = params.email + summary['E-mail on failure'] = params.email_on_fail + summary['MultiQC maxsize'] = params.max_multiqc_email_size } log.info summary.collect { k,v -> "${k.padRight(18)}: $v" }.join("\n") -log.info "\033[2m----------------------------------------------------\033[0m" +log.info "-\033[2m--------------------------------------------------\033[0m-" // Check the hostnames against configured profiles checkHostname() -def create_workflow_summary(summary) { - def yaml_file = workDir.resolve('workflow_summary_mqc.yaml') - yaml_file.text = """ +Channel.from(summary.collect{ [it.key, it.value] }) + .map { k,v -> "

$k
${v ?: 'N/A'}
" } + .reduce { a, b -> return [a, b].join("\n ") } + .map { x -> """ id: 'nf-core-atacseq-summary' description: " - this information is collected when the pipeline is started." section_name: 'nf-core/atacseq Workflow Summary' @@ -166,26 +165,23 @@ def create_workflow_summary(summary) { plot_type: 'html' data: |
-${summary.collect { k,v -> "
$k
${v ?: 'N/A'}
" }.join("\n")} + $x
- """.stripIndent() - - return yaml_file -} - + """.stripIndent() } + .set { ch_workflow_summary } /* * Parse software version numbers */ process get_software_versions { publishDir "${params.outdir}/pipeline_info", mode: 'copy', - saveAs: {filename -> - if (filename.indexOf(".csv") > 0) filename - else null - } + saveAs: { filename -> + if (filename.indexOf(".csv") > 0) filename + else null + } output: - file 'software_versions_mqc.yaml' into software_versions_yaml + file 'software_versions_mqc.yaml' into ch_software_versions_yaml file "software_versions.csv" script: @@ -199,30 +195,29 @@ process get_software_versions { """ } - - /* * STEP 1 - FastQC */ process fastqc { tag "$name" + label 'process_medium' publishDir "${params.outdir}/fastqc", mode: 'copy', - saveAs: {filename -> filename.indexOf(".zip") > 0 ? "zips/$filename" : "$filename"} + saveAs: { filename -> + filename.indexOf(".zip") > 0 ? "zips/$filename" : "$filename" + } input: - set val(name), file(reads) from read_files_fastqc + set val(name), file(reads) from ch_read_files_fastqc output: - file "*_fastqc.{zip,html}" into fastqc_results + file "*_fastqc.{zip,html}" into ch_fastqc_results script: """ - fastqc -q $reads + fastqc --quiet --threads $task.cpus $reads """ } - - /* * STEP 2 - MultiQC */ @@ -230,28 +225,28 @@ process multiqc { publishDir "${params.outdir}/MultiQC", mode: 'copy' input: - file multiqc_config from ch_multiqc_config + file (multiqc_config) from ch_multiqc_config + file (mqc_custom_config) from ch_multiqc_custom_config.collect().ifEmpty([]) // TODO nf-core: Add in log files from your new processes for MultiQC to find! - file ('fastqc/*') from fastqc_results.collect().ifEmpty([]) - file ('software_versions/*') from software_versions_yaml.collect() - file workflow_summary from create_workflow_summary(summary) + file ('fastqc/*') from ch_fastqc_results.collect().ifEmpty([]) + file ('software_versions/*') from ch_software_versions_yaml.collect() + file workflow_summary from ch_workflow_summary.collectFile(name: "workflow_summary_mqc.yaml") output: - file "*multiqc_report.html" into multiqc_report + file "*multiqc_report.html" into ch_multiqc_report file "*_data" file "multiqc_plots" script: rtitle = custom_runName ? "--title \"$custom_runName\"" : '' rfilename = custom_runName ? "--filename " + custom_runName.replaceAll('\\W','_').replaceAll('_+','_') + "_multiqc_report" : '' + custom_config_file = params.multiqc_config ? "--config $mqc_custom_config" : '' // TODO nf-core: Specify which MultiQC modules to use with -m for a faster run time """ - multiqc -f $rtitle $rfilename --config $multiqc_config . + multiqc -f $rtitle $rfilename $custom_config_file . """ } - - /* * STEP 3 - Output Description HTML */ @@ -266,12 +261,10 @@ process output_documentation { script: """ - markdown_to_html.r $output_docs results_description.html + markdown_to_html.py $output_docs -o results_description.html """ } - - /* * Completion e-mail notification */ @@ -279,8 +272,8 @@ workflow.onComplete { // Set up the e-mail variables def subject = "[nf-core/atacseq] Successful: $workflow.runName" - if(!workflow.success){ - subject = "[nf-core/atacseq] FAILED: $workflow.runName" + if (!workflow.success) { + subject = "[nf-core/atacseq] FAILED: $workflow.runName" } def email_fields = [:] email_fields['version'] = workflow.manifest.version @@ -298,21 +291,20 @@ workflow.onComplete { email_fields['summary']['Date Completed'] = workflow.complete email_fields['summary']['Pipeline script file path'] = workflow.scriptFile email_fields['summary']['Pipeline script hash ID'] = workflow.scriptId - if(workflow.repository) email_fields['summary']['Pipeline repository Git URL'] = workflow.repository - if(workflow.commitId) email_fields['summary']['Pipeline repository Git Commit'] = workflow.commitId - if(workflow.revision) email_fields['summary']['Pipeline Git branch/tag'] = workflow.revision - if(workflow.container) email_fields['summary']['Docker image'] = workflow.container + if (workflow.repository) email_fields['summary']['Pipeline repository Git URL'] = workflow.repository + if (workflow.commitId) email_fields['summary']['Pipeline repository Git Commit'] = workflow.commitId + if (workflow.revision) email_fields['summary']['Pipeline Git branch/tag'] = workflow.revision email_fields['summary']['Nextflow Version'] = workflow.nextflow.version email_fields['summary']['Nextflow Build'] = workflow.nextflow.build email_fields['summary']['Nextflow Compile Timestamp'] = workflow.nextflow.timestamp - // TODO nf-core: If not using MultiQC, strip out this code (including params.maxMultiqcEmailFileSize) + // TODO nf-core: If not using MultiQC, strip out this code (including params.max_multiqc_email_size) // On success try attach the multiqc report def mqc_report = null try { if (workflow.success) { - mqc_report = multiqc_report.getVal() - if (mqc_report.getClass() == ArrayList){ + mqc_report = ch_multiqc_report.getVal() + if (mqc_report.getClass() == ArrayList) { log.warn "[nf-core/atacseq] Found multiple reports from process 'multiqc', will use only one" mqc_report = mqc_report[0] } @@ -321,6 +313,12 @@ workflow.onComplete { log.warn "[nf-core/atacseq] Could not attach MultiQC report to summary email" } + // Check if we are only sending emails on failure + email_address = params.email + if (!params.email && params.email_on_fail && !workflow.success) { + email_address = params.email_on_fail + } + // Render the TXT template def engine = new groovy.text.GStringTemplateEngine() def tf = new File("$baseDir/assets/email_template.txt") @@ -333,89 +331,89 @@ workflow.onComplete { def email_html = html_template.toString() // Render the sendmail template - def smail_fields = [ email: params.email, subject: subject, email_txt: email_txt, email_html: email_html, baseDir: "$baseDir", mqcFile: mqc_report, mqcMaxSize: params.maxMultiqcEmailFileSize.toBytes() ] + def smail_fields = [ email: email_address, subject: subject, email_txt: email_txt, email_html: email_html, baseDir: "$baseDir", mqcFile: mqc_report, mqcMaxSize: params.max_multiqc_email_size.toBytes() ] def sf = new File("$baseDir/assets/sendmail_template.txt") def sendmail_template = engine.createTemplate(sf).make(smail_fields) def sendmail_html = sendmail_template.toString() // Send the HTML e-mail - if (params.email) { + if (email_address) { try { - if( params.plaintext_email ){ throw GroovyException('Send plaintext e-mail, not HTML') } - // Try to send HTML e-mail using sendmail - [ 'sendmail', '-t' ].execute() << sendmail_html - log.info "[nf-core/atacseq] Sent summary e-mail to $params.email (sendmail)" + if (params.plaintext_email) { throw GroovyException('Send plaintext e-mail, not HTML') } + // Try to send HTML e-mail using sendmail + [ 'sendmail', '-t' ].execute() << sendmail_html + log.info "[nf-core/atacseq] Sent summary e-mail to $email_address (sendmail)" } catch (all) { - // Catch failures and try with plaintext - [ 'mail', '-s', subject, params.email ].execute() << email_txt - log.info "[nf-core/atacseq] Sent summary e-mail to $params.email (mail)" + // Catch failures and try with plaintext + [ 'mail', '-s', subject, email_address ].execute() << email_txt + log.info "[nf-core/atacseq] Sent summary e-mail to $email_address (mail)" } } // Write summary e-mail HTML to a file - def output_d = new File( "${params.outdir}/pipeline_info/" ) - if( !output_d.exists() ) { - output_d.mkdirs() + def output_d = new File("${params.outdir}/pipeline_info/") + if (!output_d.exists()) { + output_d.mkdirs() } - def output_hf = new File( output_d, "pipeline_report.html" ) + def output_hf = new File(output_d, "pipeline_report.html") output_hf.withWriter { w -> w << email_html } - def output_tf = new File( output_d, "pipeline_report.txt" ) + def output_tf = new File(output_d, "pipeline_report.txt") output_tf.withWriter { w -> w << email_txt } - c_reset = params.monochrome_logs ? '' : "\033[0m"; - c_purple = params.monochrome_logs ? '' : "\033[0;35m"; c_green = params.monochrome_logs ? '' : "\033[0;32m"; + c_purple = params.monochrome_logs ? '' : "\033[0;35m"; c_red = params.monochrome_logs ? '' : "\033[0;31m"; + c_reset = params.monochrome_logs ? '' : "\033[0m"; - if (workflow.stats.ignoredCountFmt > 0 && workflow.success) { - log.info "${c_purple}Warning, pipeline completed, but with errored process(es) ${c_reset}" - log.info "${c_red}Number of ignored errored process(es) : ${workflow.stats.ignoredCountFmt} ${c_reset}" - log.info "${c_green}Number of successfully ran process(es) : ${workflow.stats.succeedCountFmt} ${c_reset}" + if (workflow.stats.ignoredCount > 0 && workflow.success) { + log.info "-${c_purple}Warning, pipeline completed, but with errored process(es) ${c_reset}-" + log.info "-${c_red}Number of ignored errored process(es) : ${workflow.stats.ignoredCount} ${c_reset}-" + log.info "-${c_green}Number of successfully ran process(es) : ${workflow.stats.succeedCount} ${c_reset}-" } - if(workflow.success){ - log.info "${c_purple}[nf-core/atacseq]${c_green} Pipeline completed successfully${c_reset}" + if (workflow.success) { + log.info "-${c_purple}[nf-core/atacseq]${c_green} Pipeline completed successfully${c_reset}-" } else { checkHostname() - log.info "${c_purple}[nf-core/atacseq]${c_red} Pipeline completed with errors${c_reset}" + log.info "-${c_purple}[nf-core/atacseq]${c_red} Pipeline completed with errors${c_reset}-" } } -def nfcoreHeader(){ +def nfcoreHeader() { // Log colors ANSI codes - c_reset = params.monochrome_logs ? '' : "\033[0m"; - c_dim = params.monochrome_logs ? '' : "\033[2m"; c_black = params.monochrome_logs ? '' : "\033[0;30m"; - c_green = params.monochrome_logs ? '' : "\033[0;32m"; - c_yellow = params.monochrome_logs ? '' : "\033[0;33m"; c_blue = params.monochrome_logs ? '' : "\033[0;34m"; - c_purple = params.monochrome_logs ? '' : "\033[0;35m"; c_cyan = params.monochrome_logs ? '' : "\033[0;36m"; + c_dim = params.monochrome_logs ? '' : "\033[2m"; + c_green = params.monochrome_logs ? '' : "\033[0;32m"; + c_purple = params.monochrome_logs ? '' : "\033[0;35m"; + c_reset = params.monochrome_logs ? '' : "\033[0m"; c_white = params.monochrome_logs ? '' : "\033[0;37m"; + c_yellow = params.monochrome_logs ? '' : "\033[0;33m"; - return """ ${c_dim}----------------------------------------------------${c_reset} + return """ -${c_dim}--------------------------------------------------${c_reset}- ${c_green},--.${c_black}/${c_green},-.${c_reset} ${c_blue} ___ __ __ __ ___ ${c_green}/,-._.--~\'${c_reset} ${c_blue} |\\ | |__ __ / ` / \\ |__) |__ ${c_yellow}} {${c_reset} ${c_blue} | \\| | \\__, \\__/ | \\ |___ ${c_green}\\`-._,-`-,${c_reset} ${c_green}`._,._,\'${c_reset} ${c_purple} nf-core/atacseq v${workflow.manifest.version}${c_reset} - ${c_dim}----------------------------------------------------${c_reset} + -${c_dim}--------------------------------------------------${c_reset}- """.stripIndent() } -def checkHostname(){ +def checkHostname() { def c_reset = params.monochrome_logs ? '' : "\033[0m" def c_white = params.monochrome_logs ? '' : "\033[0;37m" def c_red = params.monochrome_logs ? '' : "\033[1;91m" def c_yellow_bold = params.monochrome_logs ? '' : "\033[1;93m" - if(params.hostnames){ + if (params.hostnames) { def hostname = "hostname".execute().text.trim() params.hostnames.each { prof, hnames -> hnames.each { hname -> - if(hostname.contains(hname) && !workflow.profile.contains(prof)){ + if (hostname.contains(hname) && !workflow.profile.contains(prof)) { log.error "====================================================\n" + " ${c_red}WARNING!${c_reset} You are running with `-profile $workflow.profile`\n" + " but your machine hostname is ${c_white}'$hostname'${c_reset}\n" + diff --git a/nextflow.config b/nextflow.config index 050b5d8f..eba1de20 100644 --- a/nextflow.config +++ b/nextflow.config @@ -10,29 +10,35 @@ params { // Workflow flags // TODO nf-core: Specify your pipeline's command line flags + genome = false reads = "data/*{1,2}.fastq.gz" - singleEnd = false + single_end = false outdir = './results' // Boilerplate options name = false - multiqc_config = "$baseDir/assets/multiqc_config.yaml" + multiqc_config = false email = false - maxMultiqcEmailFileSize = 25.MB + email_on_fail = false + max_multiqc_email_size = 25.MB plaintext_email = false monochrome_logs = false help = false - igenomes_base = "./iGenomes" + igenomes_base = 's3://ngi-igenomes/igenomes/' tracedir = "${params.outdir}/pipeline_info" - awsqueue = false - awsregion = 'eu-west-1' - igenomesIgnore = false + igenomes_ignore = false custom_config_version = 'master' custom_config_base = "https://raw.githubusercontent.com/nf-core/configs/${params.custom_config_version}" hostnames = false config_profile_description = false config_profile_contact = false config_profile_url = false + + // Defaults only, expecting to be overwritten + max_memory = 128.GB + max_cpus = 16 + max_time = 240.h + } // Container slug. Stable releases should specify release tag! @@ -50,19 +56,33 @@ try { } profiles { - awsbatch { includeConfig 'conf/awsbatch.config' } conda { process.conda = "$baseDir/environment.yml" } debug { process.beforeScript = 'echo $HOSTNAME' } - docker { docker.enabled = true } - singularity { singularity.enabled = true } + docker { + docker.enabled = true + // Avoid this error: + // WARNING: Your kernel does not support swap limit capabilities or the cgroup is not mounted. Memory limited without swap. + // Testing this in nf-core after discussion here https://github.com/nf-core/tools/pull/351 + // once this is established and works well, nextflow might implement this behavior as new default. + docker.runOptions = '-u \$(id -u):\$(id -g)' + } + singularity { + singularity.enabled = true + singularity.autoMounts = true + } test { includeConfig 'conf/test.config' } } // Load igenomes.config if required -if(!params.igenomesIgnore){ +if (!params.igenomes_ignore) { includeConfig 'conf/igenomes.config' } +// Export this variable to prevent local Python libraries from conflicting with those in the container +env { + PYTHONNOUSERSITE = 1 +} + // Capture exit codes from upstream processes when piping process.shell = ['/bin/bash', '-euo', 'pipefail'] @@ -89,16 +109,16 @@ manifest { homePage = 'https://github.com/nf-core/atacseq' description = 'ATACSeq peak-calling and differential analysis pipeline.' mainScript = 'main.nf' - nextflowVersion = '>=0.32.0' - version = '1.0.0' + nextflowVersion = '>=19.10.0' + version = '1.1.1dev' } // Function to ensure that resource requirements don't go beyond // a maximum limit def check_max(obj, type) { - if(type == 'memory'){ + if (type == 'memory') { try { - if(obj.compareTo(params.max_memory as nextflow.util.MemoryUnit) == 1) + if (obj.compareTo(params.max_memory as nextflow.util.MemoryUnit) == 1) return params.max_memory as nextflow.util.MemoryUnit else return obj @@ -106,9 +126,9 @@ def check_max(obj, type) { println " ### ERROR ### Max memory '${params.max_memory}' is not valid! Using default value: $obj" return obj } - } else if(type == 'time'){ + } else if (type == 'time') { try { - if(obj.compareTo(params.max_time as nextflow.util.Duration) == 1) + if (obj.compareTo(params.max_time as nextflow.util.Duration) == 1) return params.max_time as nextflow.util.Duration else return obj @@ -116,7 +136,7 @@ def check_max(obj, type) { println " ### ERROR ### Max time '${params.max_time}' is not valid! Using default value: $obj" return obj } - } else if(type == 'cpus'){ + } else if (type == 'cpus') { try { return Math.min( obj, params.max_cpus as int ) } catch (all) { From f85dca520cb035cd346132ce749549abd838e784 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 21 Feb 2020 16:22:26 +0000 Subject: [PATCH 095/226] Resolve conflicts --- CHANGELOG.md | 1 + main.nf | 254 +++------------------------------------------------ 2 files changed, 15 insertions(+), 240 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 1691b0db..a48bb084 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -12,6 +12,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * [#76](https://github.com/nf-core/atacseq/issues/76) - featureCounts coupled to DESeq2 * [#80](https://github.com/nf-core/atacseq/pull/80) - Added social preview image * Added GitHub Actions CI tests +* Update template to tools `1.9` ### `Fixed` diff --git a/main.nf b/main.nf index 88b60b10..43800440 100644 --- a/main.nf +++ b/main.nf @@ -140,6 +140,8 @@ if (!(workflow.runName ==~ /[a-z]+_[a-z]+/)) { //////////////////////////////////////////////////// // Pipeline config +ch_multiqc_config = file("$baseDir/assets/multiqc_config.yaml", checkIfExists: true) +ch_multiqc_custom_config = params.multiqc_config ? Channel.fromPath(params.multiqc_config, checkIfExists: true) : Channel.empty() ch_output_docs = file("$baseDir/docs/output.md", checkIfExists: true) // JSON files required by BAMTools for alignment filtering @@ -150,7 +152,6 @@ if (params.single_end) { } // Header files for MultiQC -ch_multiqc_config = file(params.multiqc_config, checkIfExists: true) ch_mlib_peak_count_header = file("$baseDir/assets/multiqc/mlib_peak_count_header.txt", checkIfExists: true) ch_mlib_frip_score_header = file("$baseDir/assets/multiqc/mlib_frip_score_header.txt", checkIfExists: true) ch_mlib_peak_annotation_header = file("$baseDir/assets/multiqc/mlib_peak_annotation_header.txt", checkIfExists: true) @@ -1258,81 +1259,6 @@ process MergedLibConsensusPeakSetAnnotate { output: file "*.annotatePeaks.txt" into ch_mlib_macs_consensus_annotate -<<<<<<< HEAD - Mandatory arguments: - --reads [file] Path to input data (must be surrounded with quotes) - -profile [str] Configuration profile to use. Can use multiple (comma separated) - Available: conda, docker, singularity, test, awsbatch, and more - - Options: - --genome [str] Name of iGenomes reference - --single_end [bool] Specifies that the input is single-end reads - - References If not specified in the configuration file or you wish to overwrite any of the references - --fasta [file] Path to fasta reference - - Other options: - --outdir [file] The output directory where the results will be saved - --email [email] Set this parameter to your e-mail address to get a summary e-mail with details of the run sent to you when the workflow exits - --email_on_fail [email] Same as --email, except only send mail if the workflow is not successful - --max_multiqc_email_size [str] Theshold size for MultiQC report to be attached in notification email. If file generated by pipeline exceeds the threshold, it will not be attached (Default: 25MB) - -name [str] Name for the pipeline run. If not specified, Nextflow will automatically generate a random mnemonic - - AWSBatch options: - --awsqueue [str] The AWSBatch JobQueue that needs to be set when running on AWSBatch - --awsregion [str] The AWS Region for your AWS Batch job to run on - --awscli [str] Path to the AWS CLI tool - """.stripIndent() -} - -// Show help message -if (params.help) { - helpMessage() - exit 0 -} - -/* - * SET UP CONFIGURATION VARIABLES - */ - -// Check if genome exists in the config file -if (params.genomes && params.genome && !params.genomes.containsKey(params.genome)) { - exit 1, "The provided genome '${params.genome}' is not available in the iGenomes file. Currently the available genomes are ${params.genomes.keySet().join(", ")}" -} - -// TODO nf-core: Add any reference files that are needed -// Configurable reference genomes -// -// NOTE - THIS IS NOT USED IN THIS PIPELINE, EXAMPLE ONLY -// If you want to use the channel below in a process, define the following: -// input: -// file fasta from ch_fasta -// -params.fasta = params.genome ? params.genomes[ params.genome ].fasta ?: false : false -if (params.fasta) { ch_fasta = file(params.fasta, checkIfExists: true) } - -// Has the run name been specified by the user? -// this has the bonus effect of catching both -name and --name -custom_runName = params.name -if (!(workflow.runName ==~ /[a-z]+_[a-z]+/)) { - custom_runName = workflow.runName -} - -if (workflow.profile.contains('awsbatch')) { - // AWSBatch sanity checking - if (!params.awsqueue || !params.awsregion) exit 1, "Specify correct --awsqueue and --awsregion parameters on AWSBatch!" - // Check outdir paths to be S3 buckets if running on AWSBatch - // related: https://github.com/nextflow-io/nextflow/issues/813 - if (!params.outdir.startsWith('s3:')) exit 1, "Outdir not on S3 - specify S3 Bucket to run on AWSBatch!" - // Prevent trace files to be stored on S3 since S3 does not support rolling files. - if (params.tracedir.startsWith('s3:')) exit 1, "Specify a local tracedir or run without trace! S3 cannot be used for tracefiles." -} - -// Stage config files -ch_multiqc_config = file("$baseDir/assets/multiqc_config.yaml", checkIfExists: true) -ch_multiqc_custom_config = params.multiqc_config ? Channel.fromPath(params.multiqc_config, checkIfExists: true) : Channel.empty() -ch_output_docs = file("$baseDir/docs/output.md", checkIfExists: true) -======= script: prefix = "consensus_peaks.mLb.clN" """ @@ -1494,66 +1420,10 @@ process MergedLibAtaqvMkarv { /* -- -- */ /////////////////////////////////////////////////////////////////////////////// /////////////////////////////////////////////////////////////////////////////// ->>>>>>> dev /* * STEP 7 Merge library BAM files across all replicates */ -<<<<<<< HEAD -if (params.readPaths) { - if (params.single_end) { - Channel - .from(params.readPaths) - .map { row -> [ row[0], [ file(row[1][0], checkIfExists: true) ] ] } - .ifEmpty { exit 1, "params.readPaths was empty - no input files supplied" } - .into { ch_read_files_fastqc; ch_read_files_trimming } - } else { - Channel - .from(params.readPaths) - .map { row -> [ row[0], [ file(row[1][0], checkIfExists: true), file(row[1][1], checkIfExists: true) ] ] } - .ifEmpty { exit 1, "params.readPaths was empty - no input files supplied" } - .into { ch_read_files_fastqc; ch_read_files_trimming } - } -} else { - Channel - .fromFilePairs(params.reads, size: params.single_end ? 1 : 2) - .ifEmpty { exit 1, "Cannot find any reads matching: ${params.reads}\nNB: Path needs to be enclosed in quotes!\nIf this is single-end data, please specify --single_end on the command line." } - .into { ch_read_files_fastqc; ch_read_files_trimming } -} - -// Header log info -log.info nfcoreHeader() -def summary = [:] -if (workflow.revision) summary['Pipeline Release'] = workflow.revision -summary['Run Name'] = custom_runName ?: workflow.runName -// TODO nf-core: Report custom parameters here -summary['Reads'] = params.reads -summary['Fasta Ref'] = params.fasta -summary['Data Type'] = params.single_end ? 'Single-End' : 'Paired-End' -summary['Max Resources'] = "$params.max_memory memory, $params.max_cpus cpus, $params.max_time time per job" -if (workflow.containerEngine) summary['Container'] = "$workflow.containerEngine - $workflow.container" -summary['Output dir'] = params.outdir -summary['Launch dir'] = workflow.launchDir -summary['Working dir'] = workflow.workDir -summary['Script dir'] = workflow.projectDir -summary['User'] = workflow.userName -if (workflow.profile.contains('awsbatch')) { - summary['AWS Region'] = params.awsregion - summary['AWS Queue'] = params.awsqueue - summary['AWS CLI'] = params.awscli -} -summary['Config Profile'] = workflow.profile -if (params.config_profile_description) summary['Config Description'] = params.config_profile_description -if (params.config_profile_contact) summary['Config Contact'] = params.config_profile_contact -if (params.config_profile_url) summary['Config URL'] = params.config_profile_url -if (params.email || params.email_on_fail) { - summary['E-mail Address'] = params.email - summary['E-mail on failure'] = params.email_on_fail - summary['MultiQC maxsize'] = params.max_multiqc_email_size -} -log.info summary.collect { k,v -> "${k.padRight(18)}: $v" }.join("\n") -log.info "-\033[2m--------------------------------------------------\033[0m-" -======= ch_mlib_rm_orphan_bam_mrep .map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] } .groupTuple(by: [0]) @@ -1652,28 +1522,10 @@ process MergedRepBigWig { else if (filename.endsWith(".bigWig")) "$filename" else null } ->>>>>>> dev when: !params.skip_merge_replicates && replicatesExist -<<<<<<< HEAD -Channel.from(summary.collect{ [it.key, it.value] }) - .map { k,v -> "
$k
${v ?: 'N/A'}
" } - .reduce { a, b -> return [a, b].join("\n ") } - .map { x -> """ - id: 'nf-core-atacseq-summary' - description: " - this information is collected when the pipeline is started." - section_name: 'nf-core/atacseq Workflow Summary' - section_href: 'https://github.com/nf-core/atacseq' - plot_type: 'html' - data: | -
- $x -
- """.stripIndent() } - .set { ch_workflow_summary } -======= input: set val(name), file(bam), file(flagstat) from ch_mrep_bam_bigwig.join(ch_mrep_bam_flagstat_bigwig, by: [0]) file sizes from ch_genome_sizes_mrep_bigwig.collect() @@ -1751,23 +1603,10 @@ process MergedRepMACSCallPeak { find * -type f -name "*.${PEAK_TYPE}" -exec echo -e "bwa/mergedReplicate/macs/${PEAK_TYPE}/"{}"\\t0,0,178" \\; > ${prefix}_peaks.igv.txt """ } ->>>>>>> dev /* * STEP 8.3 Annotate peaks with HOMER */ -<<<<<<< HEAD -process get_software_versions { - publishDir "${params.outdir}/pipeline_info", mode: 'copy', - saveAs: { filename -> - if (filename.indexOf(".csv") > 0) filename - else null - } - - output: - file 'software_versions_mqc.yaml' into ch_software_versions_yaml - file "software_versions.csv" -======= process MergedRepAnnotatePeaks { tag "$name" label "process_medium" @@ -1783,7 +1622,6 @@ process MergedRepAnnotatePeaks { output: file "*.txt" into ch_mrep_macs_annotate ->>>>>>> dev script: prefix = "${name}.mRp.clN" @@ -1798,8 +1636,6 @@ process MergedRepAnnotatePeaks { """ } -<<<<<<< HEAD -======= /* * STEP 8.4 Aggregated QC plots for peaks, FRiP and peak-to-gene annotation */ @@ -1838,32 +1674,9 @@ process MergedRepPeakQC { """ } ->>>>>>> dev /* * STEP 8.5 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection */ -<<<<<<< HEAD -process fastqc { - tag "$name" - label 'process_medium' - publishDir "${params.outdir}/fastqc", mode: 'copy', - saveAs: { filename -> - filename.indexOf(".zip") > 0 ? "zips/$filename" : "$filename" - } - - input: - set val(name), file(reads) from ch_read_files_fastqc - - output: - file "*_fastqc.{zip,html}" into ch_fastqc_results - - script: - """ - fastqc --quiet --threads $task.cpus $reads - """ -} - -======= process MergedRepConsensusPeakSet { label 'process_long' publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: 'copy', @@ -2109,25 +1922,22 @@ process get_software_versions { """ } -def create_workflow_summary(summary) { - - def yaml_file = workDir.resolve('workflow_summary_mqc.yaml') - yaml_file.text = """ - id: 'nf-core-atacseq-summary' +Channel.from(summary.collect{ [it.key, it.value] }) + .map { k,v -> "
$k
${v ?: 'N/A'}
" } + .reduce { a, b -> return [a, b].join("\n ") } + .map { x -> """ + id: 'nf-core-chipseq-summary' description: " - this information is collected when the pipeline is started." - section_name: 'nf-core/atacseq Workflow Summary' - section_href: 'https://github.com/nf-core/atacseq' + section_name: 'nf-core/chipseq Workflow Summary' + section_href: 'https://github.com/nf-core/chipseq' plot_type: 'html' data: |
-${summary.collect { k,v -> "
$k
${v ?: 'N/A'}
" }.join("\n")} + $x
- """.stripIndent() - - return yaml_file -} + """.stripIndent() } + .set { ch_workflow_summary } ->>>>>>> dev /* * STEP 10 - MultiQC */ @@ -2138,18 +1948,11 @@ process MultiQC { !params.skip_multiqc input: -<<<<<<< HEAD file (multiqc_config) from ch_multiqc_config file (mqc_custom_config) from ch_multiqc_custom_config.collect().ifEmpty([]) - // TODO nf-core: Add in log files from your new processes for MultiQC to find! - file ('fastqc/*') from ch_fastqc_results.collect().ifEmpty([]) - file ('software_versions/*') from ch_software_versions_yaml.collect() - file workflow_summary from ch_workflow_summary.collectFile(name: "workflow_summary_mqc.yaml") -======= - file multiqc_config from ch_multiqc_config file ('software_versions/*') from ch_software_versions_mqc.collect() - file ('workflow_summary/*') from create_workflow_summary(summary) + file workflow_summary from ch_workflow_summary.collectFile(name: "workflow_summary_mqc.yaml") file ('fastqc/*') from ch_fastqc_reports_mqc.collect().ifEmpty([]) file ('trimgalore/*') from ch_trimgalore_results_mqc.collect().ifEmpty([]) @@ -2177,7 +1980,6 @@ process MultiQC { file ('macs/mergedReplicate/*') from ch_mrep_peak_qc_mqc.collect().ifEmpty([]) file ('macs/mergedReplicate/consensus/*') from ch_mrep_macs_consensus_counts_mqc.collect().ifEmpty([]) file ('macs/mergedReplicate/consensus/*') from ch_mrep_macs_consensus_deseq_mqc.collect().ifEmpty([]) ->>>>>>> dev output: file "*multiqc_report.html" into ch_multiqc_report @@ -2187,17 +1989,9 @@ process MultiQC { script: rtitle = custom_runName ? "--title \"$custom_runName\"" : '' rfilename = custom_runName ? "--filename " + custom_runName.replaceAll('\\W','_').replaceAll('_+','_') + "_multiqc_report" : '' -<<<<<<< HEAD custom_config_file = params.multiqc_config ? "--config $mqc_custom_config" : '' - // TODO nf-core: Specify which MultiQC modules to use with -m for a faster run time """ - multiqc -f $rtitle $rfilename $custom_config_file . - """ -} - -======= - """ - multiqc . -f $rtitle $rfilename --config $multiqc_config \\ + multiqc . -f $rtitle $rfilename $custom_config_file \\ -m custom_content -m fastqc -m cutadapt -m samtools -m picard -m preseq -m featureCounts -m deeptools """ } @@ -2210,7 +2004,6 @@ process MultiQC { /////////////////////////////////////////////////////////////////////////////// /////////////////////////////////////////////////////////////////////////////// ->>>>>>> dev /* * STEP 11 - Output description HTML */ @@ -2258,18 +2051,10 @@ workflow.onComplete { if (workflow.repository) email_fields['summary']['Pipeline repository Git URL'] = workflow.repository if (workflow.commitId) email_fields['summary']['Pipeline repository Git Commit'] = workflow.commitId if (workflow.revision) email_fields['summary']['Pipeline Git branch/tag'] = workflow.revision -<<<<<<< HEAD -======= - if (workflow.container) email_fields['summary']['Docker image'] = workflow.container ->>>>>>> dev email_fields['summary']['Nextflow Version'] = workflow.nextflow.version email_fields['summary']['Nextflow Build'] = workflow.nextflow.build email_fields['summary']['Nextflow Compile Timestamp'] = workflow.nextflow.timestamp -<<<<<<< HEAD - // TODO nf-core: If not using MultiQC, strip out this code (including params.max_multiqc_email_size) -======= ->>>>>>> dev // On success try attach the multiqc report def mqc_report = null try { @@ -2337,23 +2122,12 @@ workflow.onComplete { c_reset = params.monochrome_logs ? '' : "\033[0m"; if (workflow.stats.ignoredCount > 0 && workflow.success) { -<<<<<<< HEAD log.info "-${c_purple}Warning, pipeline completed, but with errored process(es) ${c_reset}-" log.info "-${c_red}Number of ignored errored process(es) : ${workflow.stats.ignoredCount} ${c_reset}-" log.info "-${c_green}Number of successfully ran process(es) : ${workflow.stats.succeedCount} ${c_reset}-" } - if (workflow.success) { log.info "-${c_purple}[nf-core/atacseq]${c_green} Pipeline completed successfully${c_reset}-" -======= - log.info "${c_purple}Warning, pipeline completed, but with errored process(es) ${c_reset}" - log.info "${c_red}Number of ignored errored process(es) : ${workflow.stats.ignoredCount} ${c_reset}" - log.info "${c_green}Number of successfully ran process(es) : ${workflow.stats.succeedCount} ${c_reset}" - } - - if (workflow.success) { - log.info "${c_purple}[nf-core/atacseq]${c_green} Pipeline completed successfully${c_reset}" ->>>>>>> dev } else { checkHostname() log.info "-${c_purple}[nf-core/atacseq]${c_red} Pipeline completed with errors${c_reset}-" From a20418663542d3fabdec1b160df1cea367537cb1 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 21 Feb 2020 17:16:46 +0000 Subject: [PATCH 096/226] Downgrade markdown --- CHANGELOG.md | 2 +- environment.yml | 2 +- 2 files changed, 2 insertions(+), 2 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index a48bb084..effa777a 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -24,7 +24,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Dependencies` * Add python `3.7.6` -* Add markdown `3.2.1` +* Add markdown `3.1.1` * Add pymdown-extensions `6.0` * Add pygments `2.5.2` * Add pigz `2.3.4` diff --git a/environment.yml b/environment.yml index 1caafa81..af38f938 100755 --- a/environment.yml +++ b/environment.yml @@ -8,7 +8,7 @@ channels: dependencies: ## conda-forge packages - conda-forge::python=3.7.6 - - conda-forge::markdown=3.2.1 + - conda-forge::markdown=3.1.1 - conda-forge::pymdown-extensions=6.0 - conda-forge::pygments=2.5.2 - conda-forge::r-base=3.6.2 From acf62e1b64512f6fe1b74b246cc8660308246e5d Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sat, 22 Feb 2020 22:35:22 +0000 Subject: [PATCH 097/226] Add skip_diff_analysis --- CHANGELOG.md | 6 +----- 1 file changed, 1 insertion(+), 5 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index effa777a..4533fb7a 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -11,8 +11,8 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * [#75](https://github.com/nf-core/atacseq/issues/75) - Include gene annotation versions in multiqc report * [#76](https://github.com/nf-core/atacseq/issues/76) - featureCounts coupled to DESeq2 * [#80](https://github.com/nf-core/atacseq/pull/80) - Added social preview image -* Added GitHub Actions CI tests * Update template to tools `1.9` +* Add `--skip_consensus_peaks` to skip consensus peak generation ### `Fixed` @@ -57,10 +57,6 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Deprecated` -| Deprecated | Replacement | -|------------------------------|---------------------------| -| `--skip_diff_analysis` | `--skip_consensus_peaks` | - ## [1.1.0] - 2019-11-05 ### `Added` From 54626f0eac3de910e6452dbf87dd702e1845ab96 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sat, 22 Feb 2020 22:35:26 +0000 Subject: [PATCH 098/226] Add skip_diff_analysis --- docs/usage.md | 7 ++++++- 1 file changed, 6 insertions(+), 1 deletion(-) diff --git a/docs/usage.md b/docs/usage.md index f3c619dd..9e689f68 100755 --- a/docs/usage.md +++ b/docs/usage.md @@ -42,6 +42,7 @@ * [`--min_reps_consensus`](#--min_reps_consensus) * [`--save_macs_pileup`](#--save_macs_pileup) * [`--skip_consensus_peaks`](#--skip_consensus_peaks) + * [`--skip_diff_analysis`](#--skip_diff_analysis) * [Skipping QC steps](#skipping-qc-steps) * [Job resources](#job-resources) * [Automatic resubmission](#automatic-resubmission) @@ -396,7 +397,11 @@ Instruct MACS2 to create bedGraph files using the `-B --SPMR` parameters. ### `--skip_consensus_peaks` -Skip consensus peak generation and differential binding analysis. +Skip consensus peak generation, annotation and counting. + +### `--skip_diff_analysis` + +Skip differential accessibility analysis with DESeq2. ## Skipping QC steps From 77a49454bea3a15e30a755678e4116b56bc7aa55 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sat, 22 Feb 2020 22:35:29 +0000 Subject: [PATCH 099/226] Add skip_diff_analysis --- main.nf | 8 +++++--- 1 file changed, 5 insertions(+), 3 deletions(-) diff --git a/main.nf b/main.nf index 43800440..6eaab541 100644 --- a/main.nf +++ b/main.nf @@ -62,7 +62,8 @@ def helpMessage() { --broad_cutoff [float] Specifies broad cutoff value for MACS2. Only used when --narrow_peak isnt specified (Default: 0.1) --min_reps_consensus [int] Number of biological replicates required from a given condition for a peak to contribute to a consensus peak (Default: 1) --save_macs_pileup [bool] Instruct MACS2 to create bedGraph files normalised to signal per million reads - --skip_consensus_peaks [bool] Skip consensus peak generation and differential binding analysis + --skip_consensus_peaks [bool] Skip consensus peak generation + --skip_diff_analysis [bool] Skip differential accessibility analysis QC --skip_fastqc [bool] Skip FastQC @@ -254,6 +255,7 @@ if (params.save_align_intermeds) summary['Save Intermeds'] = 'Yes' if (params.save_macs_pileup) summary['Save MACS2 Pileup'] = 'Yes' if (params.skip_merge_replicates) summary['Skip Merge Replicates'] = 'Yes' if (params.skip_consensus_peaks) summary['Skip Consensus Peaks'] = 'Yes' +if (params.skip_diff_analysis) summary['Skip Differential Analysis'] = 'Yes' if (params.skip_fastqc) summary['Skip FastQC'] = 'Yes' if (params.skip_picard_metrics) summary['Skip Picard Metrics'] = 'Yes' if (params.skip_preseq) summary['Skip Preseq'] = 'Yes' @@ -1322,7 +1324,7 @@ process MergedLibConsensusPeakSetDESeq { } when: - params.macs_gsize && replicatesExist && multipleGroups && !params.skip_consensus_peaks + params.macs_gsize && replicatesExist && multipleGroups && !params.skip_consensus_peaks && !params.skip_diff_analysis input: file counts from ch_mlib_macs_consensus_counts @@ -1808,7 +1810,7 @@ process MergedRepConsensusPeakSetDESeq { } when: - !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks + !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks && !params.skip_diff_analysis input: file counts from ch_mrep_macs_consensus_counts From a3f9b09fcb816652244cfc8fc372218197cba5e7 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sat, 22 Feb 2020 22:35:32 +0000 Subject: [PATCH 100/226] Add skip_diff_analysis --- nextflow.config | 1 + 1 file changed, 1 insertion(+) diff --git a/nextflow.config b/nextflow.config index 2b0fc062..e5400b59 100755 --- a/nextflow.config +++ b/nextflow.config @@ -41,6 +41,7 @@ params { min_reps_consensus = 1 save_macs_pileup = false skip_consensus_peaks = false + skip_diff_analysis = false // Options: QC skip_fastqc = false From b42ca1c30dcec9869931598ee96c48aa98ee7193 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sat, 22 Feb 2020 22:54:26 +0000 Subject: [PATCH 101/226] Multi-core DESeq2 --- CHANGELOG.md | 3 ++- 1 file changed, 2 insertions(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 4533fb7a..126041aa 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -10,9 +10,10 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * [#63](https://github.com/nf-core/atacseq/issues/63) - Added multicore support for Trim Galore! * [#75](https://github.com/nf-core/atacseq/issues/75) - Include gene annotation versions in multiqc report * [#76](https://github.com/nf-core/atacseq/issues/76) - featureCounts coupled to DESeq2 +* [79](https://github.com/nf-core/atacseq/issues/79) - Parallelize DESeq2 * [#80](https://github.com/nf-core/atacseq/pull/80) - Added social preview image * Update template to tools `1.9` -* Add `--skip_consensus_peaks` to skip consensus peak generation +* Parameter `--skip_consensus_peaks` to skip consensus peak generation ### `Fixed` From d917a14433041f16f2de712cdf7599bb6f68a150 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sat, 22 Feb 2020 22:54:31 +0000 Subject: [PATCH 102/226] Multi-core DESeq2 --- bin/featurecounts_deseq2.r | 12 +++++++----- 1 file changed, 7 insertions(+), 5 deletions(-) diff --git a/bin/featurecounts_deseq2.r b/bin/featurecounts_deseq2.r index c6683cbb..d9c07a0f 100755 --- a/bin/featurecounts_deseq2.r +++ b/bin/featurecounts_deseq2.r @@ -25,6 +25,7 @@ library(ggplot2) library(RColorBrewer) library(pheatmap) library(lattice) +library(BiocParallel) ################################################ ################################################ @@ -32,11 +33,12 @@ library(lattice) ################################################ ################################################ -option_list <- list(make_option(c("-i", "--featurecount_file"), type="character", default=NULL, help="Feature count file generated by the SubRead featureCounts command.", metavar="featurecount_file"), - make_option(c("-b", "--bam_suffix"), type="character", default=NULL, help="Portion of filename after sample name in featurecount file header e.g. '.rmDup.bam' if 'DRUG_R1.rmDup.bam'", metavar="bam_suffix"), +option_list <- list(make_option(c("-i", "--featurecount_file"), type="character", default=NULL, help="Feature count file generated by the SubRead featureCounts command.", metavar="path"), + make_option(c("-b", "--bam_suffix"), type="character", default=NULL, help="Portion of filename after sample name in featurecount file header e.g. '.rmDup.bam' if 'DRUG_R1.rmDup.bam'", metavar="string"), make_option(c("-o", "--outdir"), type="character", default='./', help="Output directory", metavar="path"), - make_option(c("-p", "--outprefix"), type="character", default='differential', help="Output prefix", metavar="character"), - make_option(c("-s", "--outsuffix"), type="character", default='', help="Output suffix for comparison-level results", metavar="character")) + make_option(c("-p", "--outprefix"), type="character", default='differential', help="Output prefix", metavar="string"), + make_option(c("-s", "--outsuffix"), type="character", default='', help="Output suffix for comparison-level results", metavar="string"), + make_option(c("-c", "--cores"), type="integer", default=1, help="Number of cores", metavar="integer")) opt_parser <- OptionParser(option_list=option_list) opt <- parse_args(opt_parser) @@ -86,7 +88,7 @@ if (file.exists(DDSFile) == FALSE) { counts <- count.table[,samples.vec,drop=FALSE] coldata <- data.frame(row.names=colnames(counts),condition=groups) dds <- DESeqDataSetFromMatrix(countData = round(counts), colData = coldata, design = ~ condition) - dds <- DESeq(dds) + dds <- DESeq(dds, parallel=TRUE, BPPARAM=MulticoreParam(opt$cores)) rld <- rlog(dds) save(dds,rld,file=DDSFile) } From 101d004d92e0e6a55c37f0c8587730d13acadbef Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sat, 22 Feb 2020 22:54:34 +0000 Subject: [PATCH 103/226] Multi-core DESeq2 --- main.nf | 16 ++++++++++++++-- 1 file changed, 14 insertions(+), 2 deletions(-) diff --git a/main.nf b/main.nf index 6eaab541..9a127400 100644 --- a/main.nf +++ b/main.nf @@ -1343,7 +1343,13 @@ process MergedLibConsensusPeakSetDESeq { prefix = "consensus_peaks.mLb.clN" bam_ext = params.single_end ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" """ - featurecounts_deseq2.r -i $counts -b '$bam_ext' -o ./ -p $prefix -s .mLb + featurecounts_deseq2.r \\ + --featurecount_file $counts \\ + --bam_suffix '$bam_ext' \\ + --outdir ./ \\ + --outprefix $prefix \\ + --outsuffix .mLb \\ + --cores $task.cpus cat $mlib_deseq2_pca_header ${prefix}.pca.vals.txt > ${prefix}.pca.vals_mqc.tsv cat $mlib_deseq2_clustering_header ${prefix}.sample.dists.txt > ${prefix}.sample.dists_mqc.tsv @@ -1829,7 +1835,13 @@ process MergedRepConsensusPeakSetDESeq { prefix = "consensus_peaks.mRp.clN" bam_ext = params.single_end ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" """ - featurecounts_deseq2.r -i $counts -b '$bam_ext' -o ./ -p $prefix -s .mLb + featurecounts_deseq2.r \\ + --featurecount_file $counts \\ + --bam_suffix '$bam_ext' \\ + --outdir ./ \\ + --outprefix $prefix \\ + --outsuffix .mRp \\ + --cores $task.cpus cat $mrep_deseq2_pca_header ${prefix}.pca.vals.txt > ${prefix}.pca.vals_mqc.tsv cat $mrep_deseq2_clustering_header ${prefix}.sample.dists.txt > ${prefix}.sample.dists_mqc.tsv From c4abdf831c85b67d6794a49946ec981af991e23b Mon Sep 17 00:00:00 2001 From: drpatelh Date: Mon, 24 Feb 2020 09:38:49 +0000 Subject: [PATCH 104/226] Fix long run-time for large intersects --- bin/plot_peak_intersect.r | 4 +++- 1 file changed, 3 insertions(+), 1 deletion(-) diff --git a/bin/plot_peak_intersect.r b/bin/plot_peak_intersect.r index 2404b8b9..40abf6d3 100755 --- a/bin/plot_peak_intersect.r +++ b/bin/plot_peak_intersect.r @@ -44,11 +44,13 @@ if (file.exists(OutDir) == FALSE) { comb.dat <- read.table(opt$input_file,sep="\t",header=FALSE) comb.vec <- comb.dat[,2] comb.vec <- setNames(comb.vec,comb.dat[,1]) - sets <- sort(unique(unlist(strsplit(names(comb.vec),split='&'))), decreasing = TRUE) + nintersects = length(names(comb.vec)) if (nintersects > 70) { nintersects <- 70 + comb.vec <- sort(comb.vec, decreasing = TRUE)[1:70] + sets <- sort(unique(unlist(strsplit(names(comb.vec),split='&'))), decreasing = TRUE) } pdf(opt$output_file,onefile=F,height=10,width=20) From 09a25f69a5b4559c1b0e53f33c3670eba1993fea Mon Sep 17 00:00:00 2001 From: drpatelh Date: Mon, 24 Feb 2020 10:18:09 +0000 Subject: [PATCH 105/226] Change reference_genome outdir --- docs/output.md | 6 +++--- 1 file changed, 3 insertions(+), 3 deletions(-) diff --git a/docs/output.md b/docs/output.md index 121d5c84..59fbf36b 100755 --- a/docs/output.md +++ b/docs/output.md @@ -291,7 +291,7 @@ You can skip this portion of the analysis by specifying the `--skip_merge_replic *Description*: An IGV session file will be created at the end of the pipeline containing the normalised bigWig tracks, per-sample peaks, consensus peaks and differential sites. This avoids having to load all of the data individually into IGV for visualisation. - The genome fasta file required for the IGV session will be the same as the one that was provided to the pipeline. This will be copied into `reference_genome/` to overcome any loading issues. If you prefer to use another path or an in-built genome provided by IGV just change the `genome` entry in the second-line of the session file. + The genome fasta file required for the IGV session will be the same as the one that was provided to the pipeline. This will be copied into `genome/` to overcome any loading issues. If you prefer to use another path or an in-built genome provided by IGV just change the `genome` entry in the second-line of the session file. The file paths in the IGV session file will only work if the results are kept in the same place on your storage. If the results are moved or for example, if you prefer to load the data over the web then just replace the file paths with others that are more appropriate. @@ -317,9 +317,9 @@ You can skip this portion of the analysis by specifying the `--skip_merge_replic Reference genome-specific files can be useful to keep for the downstream processing of the results. *Output directories*: - * `reference_genome/` + * `genome/` A number of genome-specific files are generated by the pipeline in order to aid in the filtering of the data, and because they are required by standard tools such as BEDTools. These can be found in this directory along with the genome fasta file which is required by IGV. If using a genome from AWS iGenomes and if it exists a `README.txt` file containing information about the annotation version will also be saved in this directory. - * `reference_genome/BWAIndex/` + * `genome/BWAIndex/` If the `--save_reference` parameter is provided then the alignment indices generated by the pipeline will be saved in this directory. This can be quite a time-consuming process so it permits their reuse for future runs of the pipeline or for other purposes. 2. **Pipeline information** From 1237b27e7da1a9d8284b649741d2f60a149fced2 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Mon, 24 Feb 2020 10:18:20 +0000 Subject: [PATCH 106/226] Fix annotation readme copy --- main.nf | 3279 ++++++++++++++++++++++++++++--------------------------- 1 file changed, 1641 insertions(+), 1638 deletions(-) diff --git a/main.nf b/main.nf index 9a127400..12a67b31 100644 --- a/main.nf +++ b/main.nf @@ -175,7 +175,12 @@ if (params.gtf) { ch_gtf = file(params.gtf, checkIfExists: true) } else if (params.gene_bed) { ch_gene_bed = file(params.gene_bed, checkIfExists: true) } if (params.tss_bed) { ch_tss_bed = file(params.tss_bed, checkIfExists: true) } if (params.blacklist) { ch_blacklist = Channel.fromPath(params.blacklist, checkIfExists: true) } else { ch_blacklist = Channel.empty() } -if (params.anno_readme && file(params.anno_readme).exists()) { ch_anno_readme = Channel.fromPath(params.anno_readme) } else { ch_anno_readme = Channel.empty() } + +// Save AWS IGenomes file conatining annotation version +if (params.anno_readme && file(params.anno_readme).exists()) { + file("${params.outdir}/genome/").mkdirs() + file(params.anno_readme).copyTo("${params.outdir}/genome/") +} if (params.fasta) { lastPath = params.fasta.lastIndexOf(File.separator) @@ -379,7 +384,7 @@ if (!params.bwa_index) { process BWAIndex { tag "$fasta" label 'process_high' - publishDir path: { params.save_reference ? "${params.outdir}/reference_genome" : params.outdir }, + publishDir path: { params.save_reference ? "${params.outdir}/genome" : params.outdir }, saveAs: { params.save_reference ? it : null }, mode: 'copy' input: @@ -403,7 +408,7 @@ if (!params.gene_bed) { process MakeGeneBED { tag "$gtf" label 'process_low' - publishDir "${params.outdir}/reference_genome", mode: 'copy' + publishDir "${params.outdir}/genome", mode: 'copy' input: file gtf from ch_gtf @@ -424,7 +429,7 @@ if (!params.gene_bed) { if (!params.tss_bed) { process MakeTSSBED { tag "$bed" - publishDir "${params.outdir}/reference_genome", mode: 'copy' + publishDir "${params.outdir}/genome", mode: 'copy' input: file bed from ch_gene_bed @@ -444,17 +449,15 @@ if (!params.tss_bed) { */ process MakeGenomeFilter { tag "$fasta" - publishDir "${params.outdir}/reference_genome", mode: 'copy' + publishDir "${params.outdir}/genome", mode: 'copy' input: file fasta from ch_fasta file blacklist from ch_blacklist.ifEmpty([]) - file readme from ch_anno_readme.ifEmpty([]) output: file "$fasta" // FASTA FILE FOR IGV file "*.fai" // FAI INDEX FOR REFERENCE GENOME - //file "$readme" // AWS IGENOMES FILE CONTAINING ANNOTATION VERSION file "*.bed" into ch_genome_filter_regions // BED FILE WITHOUT BLACKLIST REGIONS & MITOCHONDRIAL CONTIG FOR FILTERING file "*.txt" into ch_genome_autosomes // TEXT FILE CONTAINING LISTING OF AUTOSOMAL CHROMOSOMES FOR ATAQV file "*.sizes" into ch_genome_sizes_mlib_bigwig, // CHROMOSOME SIZES FILE FOR BEDTOOLS @@ -517,1637 +520,1637 @@ process FastQC { } } -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// -/* -- -- */ -/* -- ADAPTER TRIMMING -- */ -/* -- -- */ -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// - -/* - * STEP 2 - Trim Galore! - */ -if (params.skip_trimming) { - ch_trimmed_reads = ch_raw_reads_trimgalore - ch_trimgalore_results_mqc = Channel.empty() - ch_trimgalore_fastqc_reports_mqc = Channel.empty() -} else { - process TrimGalore { - tag "$name" - label 'process_high' - publishDir "${params.outdir}/trim_galore", mode: 'copy', - saveAs: { filename -> - if (filename.endsWith(".html")) "fastqc/$filename" - else if (filename.endsWith(".zip")) "fastqc/zips/$filename" - else if (filename.endsWith("trimming_report.txt")) "logs/$filename" - else params.save_trimmed ? filename : null - } - - input: - set val(name), file(reads) from ch_raw_reads_trimgalore - - output: - set val(name), file("*.fq.gz") into ch_trimmed_reads - file "*.txt" into ch_trimgalore_results_mqc - file "*.{zip,html}" into ch_trimgalore_fastqc_reports_mqc - - script: - // Calculate number of --cores for TrimGalore based on value of task.cpus - // See: https://github.com/FelixKrueger/TrimGalore/blob/master/Changelog.md#version-060-release-on-1-mar-2019 - // See: https://github.com/nf-core/atacseq/pull/65 - def cores = 1 - if (task.cpus) { - cores = (task.cpus as int) - 4 - if (params.single_end) cores = (task.cpus as int) - 3 - if (cores < 1) cores = 1 - if (cores > 4) cores = 4 - } - - // Added soft-links to original fastqs for consistent naming in MultiQC - c_r1 = params.clip_r1 > 0 ? "--clip_r1 ${params.clip_r1}" : '' - c_r2 = params.clip_r2 > 0 ? "--clip_r2 ${params.clip_r2}" : '' - tpc_r1 = params.three_prime_clip_r1 > 0 ? "--three_prime_clip_r1 ${params.three_prime_clip_r1}" : '' - tpc_r2 = params.three_prime_clip_r2 > 0 ? "--three_prime_clip_r2 ${params.three_prime_clip_r2}" : '' - nextseq = params.trim_nextseq > 0 ? "--nextseq ${params.trim_nextseq}" : '' - - // Added soft-links to original fastqs for consistent naming in MultiQC - if (params.single_end) { - """ - [ ! -f ${name}.fastq.gz ] && ln -s $reads ${name}.fastq.gz - trim_galore --cores $cores --fastqc --gzip $c_r1 $tpc_r1 $nextseq ${name}.fastq.gz - """ - } else { - """ - [ ! -f ${name}_1.fastq.gz ] && ln -s ${reads[0]} ${name}_1.fastq.gz - [ ! -f ${name}_2.fastq.gz ] && ln -s ${reads[1]} ${name}_2.fastq.gz - trim_galore --cores $cores --paired --fastqc --gzip $c_r1 $c_r2 $tpc_r1 $tpc_r2 $nextseq ${name}_1.fastq.gz ${name}_2.fastq.gz - """ - } - } -} - -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// -/* -- -- */ -/* -- ALIGN -- */ -/* -- -- */ -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// - -/* - * STEP 3.1 - Align read 1 with bwa - */ -process BWAMem { - tag "$name" - label 'process_high' - - input: - set val(name), file(reads) from ch_trimmed_reads - file index from ch_bwa_index.collect() - - output: - set val(name), file("*.bam") into ch_bwa_bam - - script: - prefix = "${name}.Lb" - rg = "\'@RG\\tID:${name}\\tSM:${name.split('_')[0..-2].join('_')}\\tPL:ILLUMINA\\tLB:${name}\\tPU:1\'" - if (params.seq_center) { - rg = "\'@RG\\tID:${name}\\tSM:${name.split('_')[0..-2].join('_')}\\tPL:ILLUMINA\\tLB:${name}\\tPU:1\\tCN:${params.seq_center}\'" - } - """ - bwa mem \\ - -t $task.cpus \\ - -M \\ - -R $rg \\ - ${index}/${bwa_base} \\ - $reads \\ - | samtools view -@ $task.cpus -b -h -F 0x0100 -O BAM -o ${prefix}.bam - - """ -} - -/* - * STEP 3.2 - Convert .bam to coordinate sorted .bam - */ -process SortBAM { - tag "$name" - label 'process_medium' - if (params.save_align_intermeds) { - publishDir path: "${params.outdir}/bwa/library", mode: 'copy', - saveAs: { filename -> - if (filename.endsWith(".flagstat")) "samtools_stats/$filename" - else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" - else if (filename.endsWith(".stats")) "samtools_stats/$filename" - else filename - } - } - - input: - set val(name), file(bam) from ch_bwa_bam - - output: - set val(name), file("*.sorted.{bam,bam.bai}") into ch_sort_bam_merge - file "*.{flagstat,idxstats,stats}" into ch_sort_bam_flagstat_mqc - - script: - prefix = "${name}.Lb" - """ - samtools sort -@ $task.cpus -o ${prefix}.sorted.bam -T $name $bam - samtools index ${prefix}.sorted.bam - samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat - samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats - samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats - """ -} - -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// -/* -- -- */ -/* -- MERGE LIBRARY BAM -- */ -/* -- -- */ -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// - -/* - * STEP 4.1 Merge BAM files for all libraries from same replicate - */ -ch_sort_bam_merge - .map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] } - .groupTuple(by: [0]) - .map { it -> [ it[0], it[1].flatten() ] } - .set { ch_sort_bam_merge } - -process MergedLibBAM { - tag "$name" - label 'process_medium' - publishDir "${params.outdir}/bwa/mergedLibrary", mode: 'copy', - saveAs: { filename -> - if (filename.endsWith(".flagstat")) "samtools_stats/$filename" - else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" - else if (filename.endsWith(".stats")) "samtools_stats/$filename" - else if (filename.endsWith(".metrics.txt")) "picard_metrics/$filename" - else params.save_align_intermeds ? filename : null - } - - input: - set val(name), file(bams) from ch_sort_bam_merge - - output: - set val(name), file("*${prefix}.sorted.{bam,bam.bai}") into ch_mlib_bam_filter, - ch_mlib_bam_preseq, - ch_mlib_bam_ataqv - file "*.{flagstat,idxstats,stats}" into ch_mlib_bam_stats_mqc - file "*.txt" into ch_mlib_bam_metrics_mqc - - script: - prefix = "${name}.mLb.mkD" - bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() - def avail_mem = 3 - if (!task.memory) { - log.info "[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." - } else { - avail_mem = task.memory.toGiga() - } - if (bam_files.size() > 1) { - """ - picard -Xmx${avail_mem}g MergeSamFiles \\ - ${'INPUT='+bam_files.join(' INPUT=')} \\ - OUTPUT=${name}.sorted.bam \\ - SORT_ORDER=coordinate \\ - VALIDATION_STRINGENCY=LENIENT \\ - TMP_DIR=tmp - samtools index ${name}.sorted.bam - - picard -Xmx${avail_mem}g MarkDuplicates \\ - INPUT=${name}.sorted.bam \\ - OUTPUT=${prefix}.sorted.bam \\ - ASSUME_SORTED=true \\ - REMOVE_DUPLICATES=false \\ - METRICS_FILE=${prefix}.MarkDuplicates.metrics.txt \\ - VALIDATION_STRINGENCY=LENIENT \\ - TMP_DIR=tmp - - samtools index ${prefix}.sorted.bam - samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats - samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat - samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats - """ - } else { - """ - picard -Xmx${avail_mem}g MarkDuplicates \\ - INPUT=${bam_files[0]} \\ - OUTPUT=${prefix}.sorted.bam \\ - ASSUME_SORTED=true \\ - REMOVE_DUPLICATES=false \\ - METRICS_FILE=${prefix}.MarkDuplicates.metrics.txt \\ - VALIDATION_STRINGENCY=LENIENT \\ - TMP_DIR=tmp - - samtools index ${prefix}.sorted.bam - samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats - samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat - samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats - """ - } -} - -/* - * STEP 4.2 Filter BAM file at merged library-level - */ -process MergedLibBAMFilter { - tag "$name" - label 'process_medium' - publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', - saveAs: { filename -> - if (params.single_end || params.save_align_intermeds) { - if (filename.endsWith(".flagstat")) "samtools_stats/$filename" - else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" - else if (filename.endsWith(".stats")) "samtools_stats/$filename" - else if (filename.endsWith(".sorted.bam")) filename - else if (filename.endsWith(".sorted.bam.bai")) filename - else null - } - } - - input: - set val(name), file(bam) from ch_mlib_bam_filter - file bed from ch_genome_filter_regions.collect() - file bamtools_filter_config from ch_bamtools_filter_config - - output: - set val(name), file("*.{bam,bam.bai}") into ch_mlib_filter_bam - set val(name), file("*.flagstat") into ch_mlib_filter_bam_flagstat - file "*.{idxstats,stats}" into ch_mlib_filter_bam_stats_mqc - - script: - prefix = params.single_end ? "${name}.mLb.clN" : "${name}.mLb.flT" - filter_params = params.single_end ? "-F 0x004" : "-F 0x004 -F 0x0008 -f 0x001" - dup_params = params.keep_dups ? "" : "-F 0x0400" - multimap_params = params.keep_multi_map ? "" : "-q 1" - blacklist_params = params.blacklist ? "-L $bed" : "" - name_sort_bam = params.single_end ? "" : "samtools sort -n -@ $task.cpus -o ${prefix}.bam -T $prefix ${prefix}.sorted.bam" - """ - samtools view \\ - $filter_params \\ - $dup_params \\ - $multimap_params \\ - $blacklist_params \\ - -b ${bam[0]} \\ - | bamtools filter \\ - -out ${prefix}.sorted.bam \\ - -script $bamtools_filter_config - - samtools index ${prefix}.sorted.bam - samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat - samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats - samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats - - $name_sort_bam - """ -} - -/* - * STEP 4.3 Remove orphan reads from paired-end BAM file - */ -if (params.single_end) { - ch_mlib_filter_bam - .into { ch_mlib_rm_orphan_bam_metrics; - ch_mlib_rm_orphan_bam_bigwig; - ch_mlib_rm_orphan_bam_macs; - ch_mlib_rm_orphan_bam_plotfingerprint; - ch_mlib_rm_orphan_bam_mrep; - ch_mlib_name_bam_mlib_counts; - ch_mlib_name_bam_mrep_counts } - - ch_mlib_filter_bam_flagstat - .into { ch_mlib_rm_orphan_flagstat_bigwig; - ch_mlib_rm_orphan_flagstat_macs; - ch_mlib_rm_orphan_flagstat_mqc } - - ch_mlib_filter_bam_stats_mqc - .set { ch_mlib_rm_orphan_stats_mqc } -} else { - process MergedLibBAMRemoveOrphan { - tag "$name" - label 'process_medium' - publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', - saveAs: { filename -> - if (filename.endsWith(".flagstat")) "samtools_stats/$filename" - else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" - else if (filename.endsWith(".stats")) "samtools_stats/$filename" - else if (filename.endsWith(".sorted.bam")) filename - else if (filename.endsWith(".sorted.bam.bai")) filename - else null - } - - input: - set val(name), file(bam) from ch_mlib_filter_bam - - output: - set val(name), file("*.sorted.{bam,bam.bai}") into ch_mlib_rm_orphan_bam_metrics, - ch_mlib_rm_orphan_bam_bigwig, - ch_mlib_rm_orphan_bam_macs, - ch_mlib_rm_orphan_bam_plotfingerprint, - ch_mlib_rm_orphan_bam_mrep - set val(name), file("${prefix}.bam") into ch_mlib_name_bam_mlib_counts, - ch_mlib_name_bam_mrep_counts - set val(name), file("*.flagstat") into ch_mlib_rm_orphan_flagstat_bigwig, - ch_mlib_rm_orphan_flagstat_macs, - ch_mlib_rm_orphan_flagstat_mqc - file "*.{idxstats,stats}" into ch_mlib_rm_orphan_stats_mqc - - script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ - prefix = "${name}.mLb.clN" - """ - bampe_rm_orphan.py ${bam[0]} ${prefix}.bam --only_fr_pairs - - samtools sort -@ $task.cpus -o ${prefix}.sorted.bam -T $prefix ${prefix}.bam - samtools index ${prefix}.sorted.bam - samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat - samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats - samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats - """ - } -} - -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// -/* -- -- */ -/* -- MERGE LIBRARY BAM POST-ANALYSIS -- */ -/* -- -- */ -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// - -/* - * STEP 5.1 preseq analysis after merging libraries and before filtering - */ -process MergedLibPreseq { - tag "$name" - label 'process_low' - publishDir "${params.outdir}/bwa/mergedLibrary/preseq", mode: 'copy' - - when: - !params.skip_preseq - - input: - set val(name), file(bam) from ch_mlib_bam_preseq - - output: - file "*.ccurve.txt" into ch_mlib_preseq_mqc - - script: - prefix = "${name}.mLb.mkD" - """ - preseq lc_extrap -v -output ${prefix}.ccurve.txt -bam ${bam[0]} - """ -} - -/* - * STEP 5.2 Picard CollectMultipleMetrics after merging libraries and filtering - */ -process MergedLibMetrics { - tag "$name" - label 'process_medium' - publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', - saveAs: { filename -> - if (filename.endsWith("_metrics")) "picard_metrics/$filename" - else if (filename.endsWith(".pdf")) "picard_metrics/pdf/$filename" - else null - } - - when: - !params.skip_picard_metrics - - input: - set val(name), file(bam) from ch_mlib_rm_orphan_bam_metrics - file fasta from ch_fasta - - output: - file "*_metrics" into ch_mlib_collectmetrics_mqc - file "*.pdf" into ch_mlib_collectmetrics_pdf - - script: - prefix = "${name}.mLb.clN" - def avail_mem = 3 - if (!task.memory) { - log.info "[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." - } else { - avail_mem = task.memory.toGiga() - } - """ - picard -Xmx${avail_mem}g CollectMultipleMetrics \\ - INPUT=${bam[0]} \\ - OUTPUT=${prefix}.CollectMultipleMetrics \\ - REFERENCE_SEQUENCE=$fasta \\ - VALIDATION_STRINGENCY=LENIENT \\ - TMP_DIR=tmp - """ -} - -/* - * STEP 5.3 Read depth normalised bigWig - */ -process MergedLibBigWig { - tag "$name" - label 'process_medium' - publishDir "${params.outdir}/bwa/mergedLibrary/bigwig", mode: 'copy', - saveAs: { filename -> - if (filename.endsWith("scale_factor.txt")) "scale/$filename" - else if (filename.endsWith(".bigWig")) "$filename" - else null - } - - input: - set val(name), file(bam), file(flagstat) from ch_mlib_rm_orphan_bam_bigwig.join(ch_mlib_rm_orphan_flagstat_bigwig, by: [0]) - file sizes from ch_genome_sizes_mlib_bigwig.collect() - - output: - set val(name), file("*.bigWig") into ch_mlib_bigwig_plotprofile - file "*scale_factor.txt" into ch_mlib_bigwig_scale - file "*igv.txt" into ch_mlib_bigwig_igv - - script: - prefix = "${name}.mLb.clN" - pe_fragment = params.single_end ? "" : "-pc" - extend = (params.single_end && params.fragment_size > 0) ? "-fs ${params.fragment_size}" : '' - """ - SCALE_FACTOR=\$(grep 'mapped (' $flagstat | awk '{print 1000000/\$1}') - echo \$SCALE_FACTOR > ${prefix}.scale_factor.txt - genomeCoverageBed -ibam ${bam[0]} -bg -scale \$SCALE_FACTOR $pe_fragment $extend | sort -T '.' -k1,1 -k2,2n > ${prefix}.bedGraph - - bedGraphToBigWig ${prefix}.bedGraph $sizes ${prefix}.bigWig - - find * -type f -name "*.bigWig" -exec echo -e "bwa/mergedLibrary/bigwig/"{}"\\t0,0,178" \\; > ${prefix}.bigWig.igv.txt - """ -} - -/* - * STEP 5.4 generate gene body coverage plot with deepTools - */ -process MergedLibPlotProfile { - tag "$name" - label 'process_high' - publishDir "${params.outdir}/bwa/mergedLibrary/deepTools/plotProfile", mode: 'copy' - - when: - !params.skip_plot_profile - - input: - set val(name), file(bigwig) from ch_mlib_bigwig_plotprofile - file bed from ch_gene_bed - - output: - file '*.{gz,pdf}' into ch_mlib_plotprofile_results - file '*.plotProfile.tab' into ch_mlib_plotprofile_mqc - - script: - prefix = "${name}.mLb.clN" - """ - computeMatrix scale-regions \\ - --regionsFileName $bed \\ - --scoreFileName $bigwig \\ - --outFileName ${prefix}.computeMatrix.mat.gz \\ - --outFileNameMatrix ${prefix}.computeMatrix.vals.mat.gz \\ - --regionBodyLength 1000 \\ - --beforeRegionStartLength 3000 \\ - --afterRegionStartLength 3000 \\ - --skipZeros \\ - --smartLabels \\ - --numberOfProcessors $task.cpus - - plotProfile --matrixFile ${prefix}.computeMatrix.mat.gz \\ - --outFileName ${prefix}.plotProfile.pdf \\ - --outFileNameData ${prefix}.plotProfile.tab - """ -} - -/* - * STEP 5.5 deepTools plotFingerprint - */ -process MergedLibPlotFingerprint { - tag "$name" - label 'process_high' - publishDir "${params.outdir}/bwa/mergedLibrary/deepTools/plotFingerprint", mode: 'copy' - - when: - !params.skip_plot_fingerprint - - input: - set val(name), file(bam) from ch_mlib_rm_orphan_bam_plotfingerprint - - output: - file '*.{txt,pdf}' into ch_mlib_plotfingerprint_results - file '*.raw.txt' into ch_mlib_plotfingerprint_mqc - - script: - prefix = "${name}.mLb.clN" - extend = (params.single_end && params.fragment_size > 0) ? "--extendReads ${params.fragment_size}" : '' - """ - plotFingerprint \\ - --bamfiles ${bam[0]} \\ - --plotFile ${prefix}.plotFingerprint.pdf \\ - $extend \\ - --labels $prefix \\ - --outRawCounts ${prefix}.plotFingerprint.raw.txt \\ - --outQualityMetrics ${prefix}.plotFingerprint.qcmetrics.txt \\ - --skipZeros \\ - --numberOfProcessors $task.cpus \\ - --numberOfSamples $params.fingerprint_bins - """ -} - -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// -/* -- -- */ -/* -- MERGE LIBRARY PEAK ANALYSIS -- */ -/* -- -- */ -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// - -/* - * STEP 6.1 Call peaks with MACS2 and calculate FRiP score - */ -process MergedLibMACSCallPeak { - tag "$name" - label 'process_medium' - publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}", mode: 'copy', - saveAs: { filename -> - if (filename.endsWith(".tsv")) "qc/$filename" - else if (filename.endsWith(".igv.txt")) null - else filename - } - - when: - params.macs_gsize - - input: - set val(name), file(bam), file(flagstat) from ch_mlib_rm_orphan_bam_macs.join(ch_mlib_rm_orphan_flagstat_macs, by: [0]) - file mlib_peak_count_header from ch_mlib_peak_count_header - file mlib_frip_score_header from ch_mlib_frip_score_header - - output: - set val(name), file("*.{bed,xls,gappedPeak,bdg}") into ch_mlib_macs_output - set val(name), file("*$PEAK_TYPE") into ch_mlib_macs_homer, - ch_mlib_macs_qc, - ch_mlib_macs_consensus, - ch_mlib_macs_ataqv - file "*igv.txt" into ch_mlib_macs_igv - file "*_mqc.tsv" into ch_mlib_macs_mqc - - script: - prefix = "${name}.mLb.clN" - broad = params.narrow_peak ? '' : "--broad --broad-cutoff ${params.broad_cutoff}" - format = params.single_end ? "BAM" : "BAMPE" - pileup = params.save_macs_pileup ? "-B --SPMR" : "" - """ - macs2 callpeak \\ - -t ${bam[0]} \\ - $broad \\ - -f $format \\ - -g $params.macs_gsize \\ - -n $prefix \\ - $pileup \\ - --keep-dup all \\ - --nomodel - - cat ${prefix}_peaks.${PEAK_TYPE} | wc -l | awk -v OFS='\t' '{ print "${name}", \$1 }' | cat $mlib_peak_count_header - > ${prefix}_peaks.count_mqc.tsv - - READS_IN_PEAKS=\$(intersectBed -a ${bam[0]} -b ${prefix}_peaks.${PEAK_TYPE} -bed -c -f 0.20 | awk -F '\t' '{sum += \$NF} END {print sum}') - grep 'mapped (' $flagstat | awk -v a="\$READS_IN_PEAKS" -v OFS='\t' '{print "${name}", a/\$1}' | cat $mlib_frip_score_header - > ${prefix}_peaks.FRiP_mqc.tsv - - find * -type f -name "*.${PEAK_TYPE}" -exec echo -e "bwa/mergedLibrary/macs/${PEAK_TYPE}/"{}"\\t0,0,178" \\; > ${prefix}_peaks.igv.txt - """ -} - -/* - * STEP 6.2 Annotate peaks with HOMER - */ -process MergedLibAnnotatePeaks { - tag "$name" - label "process_medium" - publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}", mode: 'copy' - - when: - params.macs_gsize - - input: - set val(name), file(peak) from ch_mlib_macs_homer - file fasta from ch_fasta - file gtf from ch_gtf - - output: - file "*.txt" into ch_mlib_macs_annotate - - script: - prefix = "${name}.mLb.clN" - """ - annotatePeaks.pl \\ - $peak \\ - $fasta \\ - -gid \\ - -gtf $gtf \\ - -cpu $task.cpus \\ - > ${prefix}_peaks.annotatePeaks.txt - """ -} - -/* - * STEP 6.3 Aggregated QC plots for peaks, FRiP and peak-to-gene annotation - */ -process MergedLibPeakQC { - label "process_medium" - publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/qc", mode: 'copy' - - when: - params.macs_gsize - - input: - file peaks from ch_mlib_macs_qc.collect{ it[1] } - file annos from ch_mlib_macs_annotate.collect() - file mlib_peak_annotation_header from ch_mlib_peak_annotation_header - - output: - file "*.{txt,pdf}" into ch_mlib_peak_qc - file "*.tsv" into ch_mlib_peak_qc_mqc - - script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ - suffix = 'mLb.clN' - """ - plot_macs_qc.r \\ - -i ${peaks.join(',')} \\ - -s ${peaks.join(',').replaceAll(".${suffix}_peaks.${PEAK_TYPE}","")} \\ - -o ./ \\ - -p macs_peak.${suffix} - - plot_homer_annotatepeaks.r \\ - -i ${annos.join(',')} \\ - -s ${annos.join(',').replaceAll(".${suffix}_peaks.annotatePeaks.txt","")} \\ - -o ./ \\ - -p macs_annotatePeaks.${suffix} - - cat $mlib_peak_annotation_header macs_annotatePeaks.${suffix}.summary.txt > macs_annotatePeaks.${suffix}.summary_mqc.tsv - """ -} - -/* - * STEP 6.4 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection - */ -process MergedLibConsensusPeakSet { - label 'process_long' - publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: 'copy', - saveAs: { filename -> - if (filename.endsWith(".igv.txt")) null - else filename - } - - when: - params.macs_gsize && (replicatesExist || multipleGroups) && !params.skip_consensus_peaks - - input: - file peaks from ch_mlib_macs_consensus.collect{ it[1] } - - output: - file "*.bed" into ch_mlib_macs_consensus_bed - file "*.saf" into ch_mlib_macs_consensus_saf - file "*.boolean.txt" into ch_mlib_macs_consensus_bool - file "*.intersect.{txt,plot.pdf}" into ch_mlib_macs_consensus_intersect - file "*igv.txt" into ch_mlib_macs_consensus_igv - - script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ - suffix = 'mLb.clN' - prefix = "consensus_peaks.${suffix}" - mergecols = params.narrow_peak ? (2..10).join(',') : (2..9).join(',') - collapsecols = params.narrow_peak ? (["collapse"]*9).join(',') : (["collapse"]*8).join(',') - expandparam = params.narrow_peak ? "--is_narrow_peak" : "" - """ - sort -T '.' -k1,1 -k2,2n ${peaks.collect{it.toString()}.sort().join(' ')} \\ - | mergeBed -c $mergecols -o $collapsecols > ${prefix}.txt - - macs2_merged_expand.py \\ - ${prefix}.txt \\ - ${peaks.collect{it.toString()}.sort().join(',').replaceAll("_peaks.${PEAK_TYPE}","")} \\ - ${prefix}.boolean.txt \\ - --min_replicates $params.min_reps_consensus \\ - $expandparam - - awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$1, \$2, \$3, \$4, "0", "+" }' ${prefix}.boolean.txt > ${prefix}.bed - - echo -e "GeneID\tChr\tStart\tEnd\tStrand" > ${prefix}.saf - awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$4, \$1, \$2, \$3, "+" }' ${prefix}.boolean.txt >> ${prefix}.saf - - sed -i 's/.${suffix}//g' ${prefix}.boolean.intersect.txt - plot_peak_intersect.r -i ${prefix}.boolean.intersect.txt -o ${prefix}.boolean.intersect.plot.pdf - - find * -type f -name "${prefix}.bed" -exec echo -e "bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus/"{}"\\t0,0,0" \\; > ${prefix}.bed.igv.txt - """ -} - -/* - * STEP 6.5 Annotate consensus peaks with HOMER, and add annotation to boolean output file - */ -process MergedLibConsensusPeakSetAnnotate { - label "process_medium" - publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: 'copy' - - when: - params.macs_gsize && (replicatesExist || multipleGroups) && !params.skip_consensus_peaks - - input: - file bed from ch_mlib_macs_consensus_bed - file bool from ch_mlib_macs_consensus_bool - file fasta from ch_fasta - file gtf from ch_gtf - - output: - file "*.annotatePeaks.txt" into ch_mlib_macs_consensus_annotate - - script: - prefix = "consensus_peaks.mLb.clN" - """ - annotatePeaks.pl \\ - $bed \\ - $fasta \\ - -gid \\ - -gtf $gtf \\ - -cpu $task.cpus \\ - > ${prefix}.annotatePeaks.txt - - cut -f2- ${prefix}.annotatePeaks.txt | awk 'NR==1; NR > 1 {print \$0 | "sort -T '.' -k1,1 -k2,2n"}' | cut -f6- > tmp.txt - paste $bool tmp.txt > ${prefix}.boolean.annotatePeaks.txt - """ -} - -/* - * STEP 6.6 Count reads in consensus peaks with featureCounts - */ -process MergedLibConsensusPeakSetCounts { - label 'process_medium' - publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: 'copy' - - when: - params.macs_gsize && (replicatesExist || multipleGroups) && !params.skip_consensus_peaks - - input: - file bams from ch_mlib_name_bam_mlib_counts.collect{ it[1] } - file saf from ch_mlib_macs_consensus_saf.collect() - - output: - file "*featureCounts.txt" into ch_mlib_macs_consensus_counts - file "*featureCounts.txt.summary" into ch_mlib_macs_consensus_counts_mqc - - script: - prefix = "consensus_peaks.mLb.clN" - bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() - pe_params = params.single_end ? '' : "-p --donotsort" - """ - featureCounts \\ - -F SAF \\ - -O \\ - --fracOverlap 0.2 \\ - -T $task.cpus \\ - $pe_params \\ - -a $saf \\ - -o ${prefix}.featureCounts.txt \\ - ${bam_files.join(' ')} - """ -} - -/* - * STEP 6.7 Perform differential analysis with DESeq2 - */ -process MergedLibConsensusPeakSetDESeq { - label 'process_medium' - publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus/deseq2", mode: 'copy', - saveAs: { filename -> - if (filename.endsWith(".igv.txt")) null - else filename - } - - when: - params.macs_gsize && replicatesExist && multipleGroups && !params.skip_consensus_peaks && !params.skip_diff_analysis - - input: - file counts from ch_mlib_macs_consensus_counts - file mlib_deseq2_pca_header from ch_mlib_deseq2_pca_header - file mlib_deseq2_clustering_header from ch_mlib_deseq2_clustering_header - - output: - file "*.{RData,results.txt,pdf,log}" into ch_mlib_macs_consensus_deseq_results - file "sizeFactors" into ch_mlib_macs_consensus_deseq_factors - file "*vs*/*.{pdf,txt}" into ch_mlib_macs_consensus_deseq_comp_results - file "*vs*/*.bed" into ch_mlib_macs_consensus_deseq_comp_bed - file "*igv.txt" into ch_mlib_macs_consensus_deseq_comp_igv - file "*.tsv" into ch_mlib_macs_consensus_deseq_mqc - - script: - prefix = "consensus_peaks.mLb.clN" - bam_ext = params.single_end ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" - """ - featurecounts_deseq2.r \\ - --featurecount_file $counts \\ - --bam_suffix '$bam_ext' \\ - --outdir ./ \\ - --outprefix $prefix \\ - --outsuffix .mLb \\ - --cores $task.cpus - - cat $mlib_deseq2_pca_header ${prefix}.pca.vals.txt > ${prefix}.pca.vals_mqc.tsv - cat $mlib_deseq2_clustering_header ${prefix}.sample.dists.txt > ${prefix}.sample.dists_mqc.tsv - - find * -type f -name "*.FDR0.05.results.bed" -exec echo -e "bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus/deseq2/"{}"\\t255,0,0" \\; > ${prefix}.igv.txt - """ -} - -/* - * STEP 6.8 Run ataqv on BAM file and corresponding peaks - */ -process MergedLibAtaqv { - tag "$name" - label 'process_medium' - publishDir "${params.outdir}/bwa/mergedLibrary/ataqv/${PEAK_TYPE}", mode: 'copy' - - when: - !params.skip_ataqv - - input: - set val(name), file(bam), file(peak) from ch_mlib_bam_ataqv.join(ch_mlib_macs_ataqv, by: [0]) - file autosomes from ch_genome_autosomes.collect() - file tss_bed from ch_tss_bed - - output: - file "*.json" into ch_mlib_ataqv - - script: - peak_param = params.macs_gsize ? "--peak-file ${peak}" : "" - mito_param = params.mito_name ? "--mitochondrial-reference-name ${params.mito_name}" : "" - """ - ataqv \\ - --threads $task.cpus \\ - $peak_param \\ - --tss-file $tss_bed \\ - --metrics-file ${name}.ataqv.json \\ - --name $name \\ - --ignore-read-groups \\ - --autosomal-reference-file $autosomes \\ - $mito_param \\ - NA \\ - ${bam[0]} - """ -} - -/* - * STEP 6.9 run ataqv mkarv on all JSON files to render web app - */ -process MergedLibAtaqvMkarv { - label 'process_medium' - publishDir "${params.outdir}/bwa/mergedLibrary/ataqv/${PEAK_TYPE}", mode: 'copy' - - when: - !params.skip_ataqv - - input: - file json from ch_mlib_ataqv.collect() - - output: - file "html" into ch_mlib_ataqv_mkarv - - script: - """ - mkarv \\ - --concurrency $task.cpus \\ - --force \\ - ./html/ \\ - ${json.join(' ')} - """ -} - -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// -/* -- -- */ -/* -- MERGE REPLICATE BAM -- */ -/* -- -- */ -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// - -/* - * STEP 7 Merge library BAM files across all replicates - */ -ch_mlib_rm_orphan_bam_mrep - .map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] } - .groupTuple(by: [0]) - .map { it -> [ it[0], it[1].flatten() ] } - .set { ch_mlib_rm_orphan_bam_mrep } - -process MergedRepBAM { - tag "$name" - label 'process_medium' - publishDir "${params.outdir}/bwa/mergedReplicate", mode: 'copy', - saveAs: { filename -> - if (filename.endsWith(".flagstat")) "samtools_stats/$filename" - else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" - else if (filename.endsWith(".stats")) "samtools_stats/$filename" - else if (filename.endsWith(".metrics.txt")) "picard_metrics/$filename" - else filename - } - - input: - set val(name), file(bams) from ch_mlib_rm_orphan_bam_mrep - - output: - set val(name), file("*${prefix}.sorted.{bam,bam.bai}") into ch_mrep_bam_bigwig, - ch_mrep_bam_macs - set val(name), file("*.flagstat") into ch_mrep_bam_flagstat_bigwig, - ch_mrep_bam_flagstat_macs, - ch_mrep_bam_flagstat_mqc - file "*.{idxstats,stats}" into ch_mrep_bam_stats_mqc - file "*.txt" into ch_mrep_bam_metrics_mqc - - when: - !params.skip_merge_replicates && replicatesExist - - script: - prefix = "${name}.mRp.clN" - bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() - def avail_mem = 3 - if (!task.memory) { - log.info "[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." - } else { - avail_mem = task.memory.toGiga() - } - if (bam_files.size() > 1) { - """ - picard -Xmx${avail_mem}g MergeSamFiles \\ - ${'INPUT='+bam_files.join(' INPUT=')} \\ - OUTPUT=${name}.sorted.bam \\ - SORT_ORDER=coordinate \\ - VALIDATION_STRINGENCY=LENIENT \\ - TMP_DIR=tmp - samtools index ${name}.sorted.bam - - picard -Xmx${avail_mem}g MarkDuplicates \\ - INPUT=${name}.sorted.bam \\ - OUTPUT=${prefix}.sorted.bam \\ - ASSUME_SORTED=true \\ - REMOVE_DUPLICATES=true \\ - METRICS_FILE=${prefix}.MarkDuplicates.metrics.txt \\ - VALIDATION_STRINGENCY=LENIENT \\ - TMP_DIR=tmp - - samtools index ${prefix}.sorted.bam - samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat - samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats - samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats - """ - } else { - """ - ln -s ${bams[0]} ${prefix}.sorted.bam - ln -s ${bams[1]} ${prefix}.sorted.bam.bai - touch ${prefix}.MarkDuplicates.metrics.txt - samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat - samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats - samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats - """ - } -} - -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// -/* -- -- */ -/* -- MERGE REPLICATE BAM POST-ANALYSIS -- */ -/* -- -- */ -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// - -/* - * STEP 8.1 Read depth normalised bigWig - */ -process MergedRepBigWig { - tag "$name" - label 'process_medium' - publishDir "${params.outdir}/bwa/mergedReplicate/bigwig", mode: 'copy', - saveAs: { filename -> - if (filename.endsWith("scale_factor.txt")) "scale/$filename" - else if (filename.endsWith(".bigWig")) "$filename" - else null - } - - when: - !params.skip_merge_replicates && replicatesExist - - input: - set val(name), file(bam), file(flagstat) from ch_mrep_bam_bigwig.join(ch_mrep_bam_flagstat_bigwig, by: [0]) - file sizes from ch_genome_sizes_mrep_bigwig.collect() - - output: - set val(name), file("*.bigWig") into ch_mrep_bigwig - file "*scale_factor.txt" into ch_mrep_bigwig_scale - file "*igv.txt" into ch_mrep_bigwig_igv - - script: - prefix = "${name}.mRp.clN" - pe_fragment = params.single_end ? "" : "-pc" - extend = (params.single_end && params.fragment_size > 0) ? "-fs ${params.fragment_size}" : '' - """ - SCALE_FACTOR=\$(grep 'mapped (' $flagstat | awk '{print 1000000/\$1}') - echo \$SCALE_FACTOR > ${prefix}.scale_factor.txt - genomeCoverageBed -ibam ${bam[0]} -bg -scale \$SCALE_FACTOR $pe_fragment $extend | sort -T '.' -k1,1 -k2,2n > ${prefix}.bedGraph - - bedGraphToBigWig ${prefix}.bedGraph $sizes ${prefix}.bigWig - - find * -type f -name "*.bigWig" -exec echo -e "bwa/mergedReplicate/bigwig/"{}"\\t0,0,178" \\; > ${prefix}.bigWig.igv.txt - """ -} - -/* - * STEP 8.2 Call peaks with MACS2 and calculate FRiP score - */ -process MergedRepMACSCallPeak { - tag "$name" - label 'process_medium' - publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}", mode: 'copy', - saveAs: { filename -> - if (filename.endsWith(".tsv")) "qc/$filename" - else if (filename.endsWith(".igv.txt")) null - else filename - } - - when: - !params.skip_merge_replicates && replicatesExist && params.macs_gsize - - input: - set val(name), file(bam), file(flagstat) from ch_mrep_bam_macs.join(ch_mrep_bam_flagstat_macs, by: [0]) - file mrep_peak_count_header from ch_mrep_peak_count_header - file mrep_frip_score_header from ch_mrep_frip_score_header - - output: - set val(name), file("*.{bed,xls,gappedPeak,bdg}") into ch_mrep_macs_output - set val(name), file("*$PEAK_TYPE") into ch_mrep_macs_homer, - ch_mrep_macs_qc, - ch_mrep_macs_consensus - file "*igv.txt" into ch_mrep_macs_igv - file "*_mqc.tsv" into ch_mrep_macs_mqc - - script: - prefix = "${name}.mRp.clN" - broad = params.narrow_peak ? '' : "--broad --broad-cutoff ${params.broad_cutoff}" - format = params.single_end ? "BAM" : "BAMPE" - pileup = params.save_macs_pileup ? "-B --SPMR" : "" - """ - macs2 callpeak \\ - -t ${bam[0]} \\ - $broad \\ - -f $format \\ - -g $params.macs_gsize \\ - -n $prefix \\ - $pileup \\ - --keep-dup all \\ - --nomodel - - cat ${prefix}_peaks.${PEAK_TYPE} | wc -l | awk -v OFS='\t' '{ print "${name}", \$1 }' | cat $mrep_peak_count_header - > ${prefix}_peaks.count_mqc.tsv - - READS_IN_PEAKS=\$(intersectBed -a ${bam[0]} -b ${prefix}_peaks.${PEAK_TYPE} -bed -c -f 0.20 | awk -F '\t' '{sum += \$NF} END {print sum}') - grep 'mapped (' $flagstat | awk -v a="\$READS_IN_PEAKS" -v OFS='\t' '{print "${name}", a/\$1}' | cat $mrep_frip_score_header - > ${prefix}_peaks.FRiP_mqc.tsv - - find * -type f -name "*.${PEAK_TYPE}" -exec echo -e "bwa/mergedReplicate/macs/${PEAK_TYPE}/"{}"\\t0,0,178" \\; > ${prefix}_peaks.igv.txt - """ -} - -/* - * STEP 8.3 Annotate peaks with HOMER - */ -process MergedRepAnnotatePeaks { - tag "$name" - label "process_medium" - publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}", mode: 'copy' - - when: - !params.skip_merge_replicates && replicatesExist && params.macs_gsize - - input: - set val(name), file(peak) from ch_mrep_macs_homer - file fasta from ch_fasta - file gtf from ch_gtf - - output: - file "*.txt" into ch_mrep_macs_annotate - - script: - prefix = "${name}.mRp.clN" - """ - annotatePeaks.pl \\ - $peak \\ - $fasta \\ - -gid \\ - -gtf $gtf \\ - -cpu $task.cpus \\ - > ${prefix}_peaks.annotatePeaks.txt - """ -} - -/* - * STEP 8.4 Aggregated QC plots for peaks, FRiP and peak-to-gene annotation - */ -process MergedRepPeakQC { - label "process_medium" - publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/qc", mode: 'copy' - - when: - !params.skip_merge_replicates && replicatesExist && params.macs_gsize - - input: - file peaks from ch_mrep_macs_qc.collect{ it[1] } - file annos from ch_mrep_macs_annotate.collect() - file mrep_peak_annotation_header from ch_mrep_peak_annotation_header - - output: - file "*.{txt,pdf}" into ch_mrep_peak_qc - file "*.tsv" into ch_mrep_peak_qc_mqc - - script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ - suffix = 'mRp.clN' - """ - plot_macs_qc.r \\ - -i ${peaks.join(',')} \\ - -s ${peaks.join(',').replaceAll(".${suffix}_peaks.${PEAK_TYPE}","")} \\ - -o ./ \\ - -p macs_peak.${suffix} - - plot_homer_annotatepeaks.r \\ - -i ${annos.join(',')} \\ - -s ${annos.join(',').replaceAll(".${suffix}_peaks.annotatePeaks.txt","")} \\ - -o ./ \\ - -p macs_annotatePeaks.${suffix} - - cat $mrep_peak_annotation_header macs_annotatePeaks.${suffix}.summary.txt > macs_annotatePeaks.${suffix}.summary_mqc.tsv - """ -} - -/* - * STEP 8.5 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection - */ -process MergedRepConsensusPeakSet { - label 'process_long' - publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: 'copy', - saveAs: { filename -> - if (filename.endsWith(".igv.txt")) null - else filename - } - - when: - !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks - - input: - file peaks from ch_mrep_macs_consensus.collect{ it[1] } - - output: - file "*.bed" into ch_mrep_macs_consensus_bed - file "*.saf" into ch_mrep_macs_consensus_saf - file "*.boolean.txt" into ch_mrep_macs_consensus_bool - file "*.intersect.{txt,plot.pdf}" into ch_mrep_macs_consensus_intersect - file "*igv.txt" into ch_mrep_macs_consensus_igv - - script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ - suffix = 'mRp.clN' - prefix = "consensus_peaks.${suffix}" - mergecols = params.narrow_peak ? (2..10).join(',') : (2..9).join(',') - collapsecols = params.narrow_peak ? (["collapse"]*9).join(',') : (["collapse"]*8).join(',') - expandparam = params.narrow_peak ? "--is_narrow_peak" : "" - """ - sort -T '.' -k1,1 -k2,2n ${peaks.collect{it.toString()}.sort().join(' ')} \\ - | mergeBed -c $mergecols -o $collapsecols > ${prefix}.txt - - macs2_merged_expand.py \\ - ${prefix}.txt \\ - ${peaks.collect{it.toString()}.sort().join(',').replaceAll("_peaks.${PEAK_TYPE}","")} \\ - ${prefix}.boolean.txt \\ - $expandparam - - awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$1, \$2, \$3, \$4, "0", "+" }' ${prefix}.boolean.txt > ${prefix}.bed - - echo -e "GeneID\tChr\tStart\tEnd\tStrand" > ${prefix}.saf - awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$4, \$1, \$2, \$3, "+" }' ${prefix}.boolean.txt >> ${prefix}.saf - - sed -i 's/.${suffix}//g' ${prefix}.boolean.intersect.txt - plot_peak_intersect.r -i ${prefix}.boolean.intersect.txt -o ${prefix}.boolean.intersect.plot.pdf - - find * -type f -name "${prefix}.bed" -exec echo -e "bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus/"{}"\\t0,0,0" \\; > ${prefix}.bed.igv.txt - """ -} - -/* - * STEP 8.6 Annotate consensus peaks with HOMER, and add annotation to boolean output file - */ -process MergedRepConsensusPeakSetAnnotate { - label "process_medium" - publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: 'copy' - - when: - !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks - - input: - file bed from ch_mrep_macs_consensus_bed - file bool from ch_mrep_macs_consensus_bool - file fasta from ch_fasta - file gtf from ch_gtf - - output: - file "*.annotatePeaks.txt" into ch_mrep_macs_consensus_annotate - - script: - prefix = "consensus_peaks.mRp.clN" - """ - annotatePeaks.pl \\ - $bed \\ - $fasta \\ - -gid \\ - -gtf $gtf \\ - -cpu $task.cpus \\ - > ${prefix}.annotatePeaks.txt - - cut -f2- ${prefix}.annotatePeaks.txt | awk 'NR==1; NR > 1 {print \$0 | "sort -T '.' -k1,1 -k2,2n"}' | cut -f6- > tmp.txt - paste $bool tmp.txt > ${prefix}.boolean.annotatePeaks.txt - """ -} - -/* - * STEP 8.7 Count reads in consensus peaks with featureCounts - */ -process MergedRepConsensusPeakSetCounts { - label 'process_medium' - publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: 'copy' - - when: - !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks - - input: - file bams from ch_mlib_name_bam_mrep_counts.collect{ it[1] } - file saf from ch_mrep_macs_consensus_saf.collect() - - output: - file "*featureCounts.txt" into ch_mrep_macs_consensus_counts - file "*featureCounts.txt.summary" into ch_mrep_macs_consensus_counts_mqc - - script: - prefix = "consensus_peaks.mRp.clN" - bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() - pe_params = params.single_end ? '' : "-p --donotsort" - """ - featureCounts \\ - -F SAF \\ - -O \\ - --fracOverlap 0.2 \\ - -T $task.cpus \\ - $pe_params \\ - -a $saf \\ - -o ${prefix}.featureCounts.txt \\ - ${bam_files.join(' ')} - """ -} - -/* - * STEP 8.8 Perform differential analysis with DESeq2 - */ -process MergedRepConsensusPeakSetDESeq { - label 'process_medium' - publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus/deseq2", mode: 'copy', - saveAs: { filename -> - if (filename.endsWith(".igv.txt")) null - else filename - } - - when: - !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks && !params.skip_diff_analysis - - input: - file counts from ch_mrep_macs_consensus_counts - file mrep_deseq2_pca_header from ch_mrep_deseq2_pca_header - file mrep_deseq2_clustering_header from ch_mrep_deseq2_clustering_header - - output: - file "*.{RData,results.txt,pdf,log}" into ch_mrep_macs_consensus_deseq_results - file "sizeFactors" into ch_mrep_macs_consensus_deseq_factors - file "*vs*/*.{pdf,txt}" into ch_mrep_macs_consensus_deseq_comp_results - file "*vs*/*.bed" into ch_mrep_macs_consensus_deseq_comp_bed - file "*igv.txt" into ch_mrep_macs_consensus_deseq_comp_igv - file "*.tsv" into ch_mrep_macs_consensus_deseq_mqc - - script: - prefix = "consensus_peaks.mRp.clN" - bam_ext = params.single_end ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" - """ - featurecounts_deseq2.r \\ - --featurecount_file $counts \\ - --bam_suffix '$bam_ext' \\ - --outdir ./ \\ - --outprefix $prefix \\ - --outsuffix .mRp \\ - --cores $task.cpus - - cat $mrep_deseq2_pca_header ${prefix}.pca.vals.txt > ${prefix}.pca.vals_mqc.tsv - cat $mrep_deseq2_clustering_header ${prefix}.sample.dists.txt > ${prefix}.sample.dists_mqc.tsv - - find * -type f -name "*.FDR0.05.results.bed" -exec echo -e "bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus/deseq2/"{}"\\t255,0,0" \\; > ${prefix}.igv.txt - """ -} - -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// -/* -- -- */ -/* -- IGV -- */ -/* -- -- */ -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// - -/* - * STEP 9 - Create IGV session file - */ -process IGV { - publishDir "${params.outdir}/igv/${PEAK_TYPE}", mode: 'copy' - - when: - !params.skip_igv - - input: - file fasta from ch_fasta - - file bigwigs from ch_mlib_bigwig_igv.collect().ifEmpty([]) - file peaks from ch_mlib_macs_igv.collect().ifEmpty([]) - file consensus_peaks from ch_mlib_macs_consensus_igv.collect().ifEmpty([]) - file differential_peaks from ch_mlib_macs_consensus_deseq_comp_igv.collect().ifEmpty([]) - - file rbigwigs from ch_mrep_bigwig_igv.collect().ifEmpty([]) - file rpeaks from ch_mrep_macs_igv.collect().ifEmpty([]) - file rconsensus_peaks from ch_mrep_macs_consensus_igv.collect().ifEmpty([]) - file rdifferential_peaks from ch_mrep_macs_consensus_deseq_comp_igv.collect().ifEmpty([]) - - output: - file "*.{txt,xml}" into ch_igv_session - - script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ - """ - cat *.txt > igv_files.txt - igv_files_to_session.py igv_session.xml igv_files.txt ../../reference_genome/${fasta.getName()} --path_prefix '../../' - """ -} - -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// -/* -- -- */ -/* -- MULTIQC -- */ -/* -- -- */ -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// - -/* - * Parse software version numbers - */ -process get_software_versions { - publishDir "${params.outdir}/pipeline_info", mode: 'copy', - saveAs: { filename -> - if (filename.indexOf(".csv") > 0) filename - else null - } - - output: - file 'software_versions_mqc.yaml' into ch_software_versions_mqc - file "software_versions.csv" - - script: - """ - echo $workflow.manifest.version > v_pipeline.txt - echo $workflow.nextflow.version > v_nextflow.txt - fastqc --version > v_fastqc.txt - trim_galore --version > v_trim_galore.txt - echo \$(bwa 2>&1) > v_bwa.txt - samtools --version > v_samtools.txt - bedtools --version > v_bedtools.txt - echo \$(bamtools --version 2>&1) > v_bamtools.txt - echo \$(plotFingerprint --version 2>&1) > v_deeptools.txt || true - picard MarkDuplicates --version &> v_picard.txt || true - echo \$(R --version 2>&1) > v_R.txt - python -c "import pysam; print(pysam.__version__)" > v_pysam.txt - echo \$(macs2 --version 2>&1) > v_macs2.txt - touch v_homer.txt - echo \$(ataqv --version 2>&1) > v_ataqv.txt - echo \$(featureCounts -v 2>&1) > v_featurecounts.txt - preseq &> v_preseq.txt - multiqc --version > v_multiqc.txt - scrape_software_versions.py &> software_versions_mqc.yaml - """ -} - -Channel.from(summary.collect{ [it.key, it.value] }) - .map { k,v -> "
$k
${v ?: 'N/A'}
" } - .reduce { a, b -> return [a, b].join("\n ") } - .map { x -> """ - id: 'nf-core-chipseq-summary' - description: " - this information is collected when the pipeline is started." - section_name: 'nf-core/chipseq Workflow Summary' - section_href: 'https://github.com/nf-core/chipseq' - plot_type: 'html' - data: | -
- $x -
- """.stripIndent() } - .set { ch_workflow_summary } - -/* - * STEP 10 - MultiQC - */ -process MultiQC { - publishDir "${params.outdir}/multiqc/${PEAK_TYPE}", mode: 'copy' - - when: - !params.skip_multiqc - - input: - file (multiqc_config) from ch_multiqc_config - file (mqc_custom_config) from ch_multiqc_custom_config.collect().ifEmpty([]) - - file ('software_versions/*') from ch_software_versions_mqc.collect() - file workflow_summary from ch_workflow_summary.collectFile(name: "workflow_summary_mqc.yaml") - - file ('fastqc/*') from ch_fastqc_reports_mqc.collect().ifEmpty([]) - file ('trimgalore/*') from ch_trimgalore_results_mqc.collect().ifEmpty([]) - file ('trimgalore/fastqc/*') from ch_trimgalore_fastqc_reports_mqc.collect().ifEmpty([]) - - file ('alignment/library/*') from ch_sort_bam_flagstat_mqc.collect() - - file ('alignment/mergedLibrary/*') from ch_mlib_bam_stats_mqc.collect() - file ('alignment/mergedLibrary/*') from ch_mlib_rm_orphan_flagstat_mqc.collect{it[1]} - file ('alignment/mergedLibrary/*') from ch_mlib_rm_orphan_stats_mqc.collect() - file ('alignment/mergedLibrary/picard_metrics/*') from ch_mlib_bam_metrics_mqc.collect() - file ('alignment/mergedLibrary/picard_metrics/*') from ch_mlib_collectmetrics_mqc.collect() - file ('macs/mergedLibrary/*') from ch_mlib_macs_mqc.collect().ifEmpty([]) - file ('macs/mergedLibrary/*') from ch_mlib_peak_qc_mqc.collect().ifEmpty([]) - file ('macs/mergedLibrary/consensus/*') from ch_mlib_macs_consensus_counts_mqc.collect().ifEmpty([]) - file ('macs/mergedLibrary/consensus/*') from ch_mlib_macs_consensus_deseq_mqc.collect().ifEmpty([]) - file ('preseq/*') from ch_mlib_preseq_mqc.collect().ifEmpty([]) - file ('deeptools/*') from ch_mlib_plotprofile_mqc.collect().ifEmpty([]) - file ('deeptools/*') from ch_mlib_plotfingerprint_mqc.collect().ifEmpty([]) - - file ('alignment/mergedReplicate/*') from ch_mrep_bam_flagstat_mqc.collect{it[1]}.ifEmpty([]) - file ('alignment/mergedReplicate/*') from ch_mrep_bam_stats_mqc.collect().ifEmpty([]) - file ('alignment/mergedReplicate/*') from ch_mrep_bam_metrics_mqc.collect().ifEmpty([]) - file ('macs/mergedReplicate/*') from ch_mrep_macs_mqc.collect().ifEmpty([]) - file ('macs/mergedReplicate/*') from ch_mrep_peak_qc_mqc.collect().ifEmpty([]) - file ('macs/mergedReplicate/consensus/*') from ch_mrep_macs_consensus_counts_mqc.collect().ifEmpty([]) - file ('macs/mergedReplicate/consensus/*') from ch_mrep_macs_consensus_deseq_mqc.collect().ifEmpty([]) - - output: - file "*multiqc_report.html" into ch_multiqc_report - file "*_data" - file "multiqc_plots" - - script: - rtitle = custom_runName ? "--title \"$custom_runName\"" : '' - rfilename = custom_runName ? "--filename " + custom_runName.replaceAll('\\W','_').replaceAll('_+','_') + "_multiqc_report" : '' - custom_config_file = params.multiqc_config ? "--config $mqc_custom_config" : '' - """ - multiqc . -f $rtitle $rfilename $custom_config_file \\ - -m custom_content -m fastqc -m cutadapt -m samtools -m picard -m preseq -m featureCounts -m deeptools - """ -} - -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// -/* -- -- */ -/* -- REPORTS/DOCUMENTATION -- */ -/* -- -- */ -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// - -/* - * STEP 11 - Output description HTML - */ -process output_documentation { - publishDir "${params.outdir}/Documentation", mode: 'copy' - - input: - file output_docs from ch_output_docs - - output: - file "results_description.html" - - script: - """ - markdown_to_html.py $output_docs -o results_description.html - """ -} - -/* - * Completion e-mail notification - */ -workflow.onComplete { - - // Set up the e-mail variables - def subject = "[nf-core/atacseq] Successful: $workflow.runName" - if (!workflow.success) { - subject = "[nf-core/atacseq] FAILED: $workflow.runName" - } - def email_fields = [:] - email_fields['version'] = workflow.manifest.version - email_fields['runName'] = custom_runName ?: workflow.runName - email_fields['success'] = workflow.success - email_fields['dateComplete'] = workflow.complete - email_fields['duration'] = workflow.duration - email_fields['exitStatus'] = workflow.exitStatus - email_fields['errorMessage'] = (workflow.errorMessage ?: 'None') - email_fields['errorReport'] = (workflow.errorReport ?: 'None') - email_fields['commandLine'] = workflow.commandLine - email_fields['projectDir'] = workflow.projectDir - email_fields['summary'] = summary - email_fields['summary']['Date Started'] = workflow.start - email_fields['summary']['Date Completed'] = workflow.complete - email_fields['summary']['Pipeline script file path'] = workflow.scriptFile - email_fields['summary']['Pipeline script hash ID'] = workflow.scriptId - if (workflow.repository) email_fields['summary']['Pipeline repository Git URL'] = workflow.repository - if (workflow.commitId) email_fields['summary']['Pipeline repository Git Commit'] = workflow.commitId - if (workflow.revision) email_fields['summary']['Pipeline Git branch/tag'] = workflow.revision - email_fields['summary']['Nextflow Version'] = workflow.nextflow.version - email_fields['summary']['Nextflow Build'] = workflow.nextflow.build - email_fields['summary']['Nextflow Compile Timestamp'] = workflow.nextflow.timestamp - - // On success try attach the multiqc report - def mqc_report = null - try { - if (workflow.success) { - mqc_report = ch_multiqc_report.getVal() - if (mqc_report.getClass() == ArrayList) { - log.warn "[nf-core/atacseq] Found multiple reports from process 'multiqc', will use only one" - mqc_report = mqc_report[0] - } - } - } catch (all) { - log.warn "[nf-core/atacseq] Could not attach MultiQC report to summary email" - } - - // Check if we are only sending emails on failure - email_address = params.email - if (!params.email && params.email_on_fail && !workflow.success) { - email_address = params.email_on_fail - } - - // Render the TXT template - def engine = new groovy.text.GStringTemplateEngine() - def tf = new File("$baseDir/assets/email_template.txt") - def txt_template = engine.createTemplate(tf).make(email_fields) - def email_txt = txt_template.toString() - - // Render the HTML template - def hf = new File("$baseDir/assets/email_template.html") - def html_template = engine.createTemplate(hf).make(email_fields) - def email_html = html_template.toString() - - // Render the sendmail template - def smail_fields = [ email: email_address, subject: subject, email_txt: email_txt, email_html: email_html, baseDir: "$baseDir", mqcFile: mqc_report, mqcMaxSize: params.max_multiqc_email_size.toBytes() ] - def sf = new File("$baseDir/assets/sendmail_template.txt") - def sendmail_template = engine.createTemplate(sf).make(smail_fields) - def sendmail_html = sendmail_template.toString() - - // Send the HTML e-mail - if (email_address) { - try { - if (params.plaintext_email) { throw GroovyException('Send plaintext e-mail, not HTML') } - // Try to send HTML e-mail using sendmail - [ 'sendmail', '-t' ].execute() << sendmail_html - log.info "[nf-core/atacseq] Sent summary e-mail to $email_address (sendmail)" - } catch (all) { - // Catch failures and try with plaintext - [ 'mail', '-s', subject, email_address ].execute() << email_txt - log.info "[nf-core/atacseq] Sent summary e-mail to $email_address (mail)" - } - } - - // Write summary e-mail HTML to a file - def output_d = new File("${params.outdir}/pipeline_info/") - if (!output_d.exists()) { - output_d.mkdirs() - } - def output_hf = new File(output_d, "pipeline_report.html") - output_hf.withWriter { w -> w << email_html } - def output_tf = new File(output_d, "pipeline_report.txt") - output_tf.withWriter { w -> w << email_txt } - - c_green = params.monochrome_logs ? '' : "\033[0;32m"; - c_purple = params.monochrome_logs ? '' : "\033[0;35m"; - c_red = params.monochrome_logs ? '' : "\033[0;31m"; - c_reset = params.monochrome_logs ? '' : "\033[0m"; - - if (workflow.stats.ignoredCount > 0 && workflow.success) { - log.info "-${c_purple}Warning, pipeline completed, but with errored process(es) ${c_reset}-" - log.info "-${c_red}Number of ignored errored process(es) : ${workflow.stats.ignoredCount} ${c_reset}-" - log.info "-${c_green}Number of successfully ran process(es) : ${workflow.stats.succeedCount} ${c_reset}-" - } - if (workflow.success) { - log.info "-${c_purple}[nf-core/atacseq]${c_green} Pipeline completed successfully${c_reset}-" - } else { - checkHostname() - log.info "-${c_purple}[nf-core/atacseq]${c_red} Pipeline completed with errors${c_reset}-" - } - -} +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// /* -- -- */ +// /* -- ADAPTER TRIMMING -- */ +// /* -- -- */ +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// +// /* +// * STEP 2 - Trim Galore! +// */ +// if (params.skip_trimming) { +// ch_trimmed_reads = ch_raw_reads_trimgalore +// ch_trimgalore_results_mqc = Channel.empty() +// ch_trimgalore_fastqc_reports_mqc = Channel.empty() +// } else { +// process TrimGalore { +// tag "$name" +// label 'process_high' +// publishDir "${params.outdir}/trim_galore", mode: 'copy', +// saveAs: { filename -> +// if (filename.endsWith(".html")) "fastqc/$filename" +// else if (filename.endsWith(".zip")) "fastqc/zips/$filename" +// else if (filename.endsWith("trimming_report.txt")) "logs/$filename" +// else params.save_trimmed ? filename : null +// } +// +// input: +// set val(name), file(reads) from ch_raw_reads_trimgalore +// +// output: +// set val(name), file("*.fq.gz") into ch_trimmed_reads +// file "*.txt" into ch_trimgalore_results_mqc +// file "*.{zip,html}" into ch_trimgalore_fastqc_reports_mqc +// +// script: +// // Calculate number of --cores for TrimGalore based on value of task.cpus +// // See: https://github.com/FelixKrueger/TrimGalore/blob/master/Changelog.md#version-060-release-on-1-mar-2019 +// // See: https://github.com/nf-core/atacseq/pull/65 +// def cores = 1 +// if (task.cpus) { +// cores = (task.cpus as int) - 4 +// if (params.single_end) cores = (task.cpus as int) - 3 +// if (cores < 1) cores = 1 +// if (cores > 4) cores = 4 +// } +// +// // Added soft-links to original fastqs for consistent naming in MultiQC +// c_r1 = params.clip_r1 > 0 ? "--clip_r1 ${params.clip_r1}" : '' +// c_r2 = params.clip_r2 > 0 ? "--clip_r2 ${params.clip_r2}" : '' +// tpc_r1 = params.three_prime_clip_r1 > 0 ? "--three_prime_clip_r1 ${params.three_prime_clip_r1}" : '' +// tpc_r2 = params.three_prime_clip_r2 > 0 ? "--three_prime_clip_r2 ${params.three_prime_clip_r2}" : '' +// nextseq = params.trim_nextseq > 0 ? "--nextseq ${params.trim_nextseq}" : '' +// +// // Added soft-links to original fastqs for consistent naming in MultiQC +// if (params.single_end) { +// """ +// [ ! -f ${name}.fastq.gz ] && ln -s $reads ${name}.fastq.gz +// trim_galore --cores $cores --fastqc --gzip $c_r1 $tpc_r1 $nextseq ${name}.fastq.gz +// """ +// } else { +// """ +// [ ! -f ${name}_1.fastq.gz ] && ln -s ${reads[0]} ${name}_1.fastq.gz +// [ ! -f ${name}_2.fastq.gz ] && ln -s ${reads[1]} ${name}_2.fastq.gz +// trim_galore --cores $cores --paired --fastqc --gzip $c_r1 $c_r2 $tpc_r1 $tpc_r2 $nextseq ${name}_1.fastq.gz ${name}_2.fastq.gz +// """ +// } +// } +// } +// +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// /* -- -- */ +// /* -- ALIGN -- */ +// /* -- -- */ +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// +// /* +// * STEP 3.1 - Align read 1 with bwa +// */ +// process BWAMem { +// tag "$name" +// label 'process_high' +// +// input: +// set val(name), file(reads) from ch_trimmed_reads +// file index from ch_bwa_index.collect() +// +// output: +// set val(name), file("*.bam") into ch_bwa_bam +// +// script: +// prefix = "${name}.Lb" +// rg = "\'@RG\\tID:${name}\\tSM:${name.split('_')[0..-2].join('_')}\\tPL:ILLUMINA\\tLB:${name}\\tPU:1\'" +// if (params.seq_center) { +// rg = "\'@RG\\tID:${name}\\tSM:${name.split('_')[0..-2].join('_')}\\tPL:ILLUMINA\\tLB:${name}\\tPU:1\\tCN:${params.seq_center}\'" +// } +// """ +// bwa mem \\ +// -t $task.cpus \\ +// -M \\ +// -R $rg \\ +// ${index}/${bwa_base} \\ +// $reads \\ +// | samtools view -@ $task.cpus -b -h -F 0x0100 -O BAM -o ${prefix}.bam - +// """ +// } +// +// /* +// * STEP 3.2 - Convert .bam to coordinate sorted .bam +// */ +// process SortBAM { +// tag "$name" +// label 'process_medium' +// if (params.save_align_intermeds) { +// publishDir path: "${params.outdir}/bwa/library", mode: 'copy', +// saveAs: { filename -> +// if (filename.endsWith(".flagstat")) "samtools_stats/$filename" +// else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" +// else if (filename.endsWith(".stats")) "samtools_stats/$filename" +// else filename +// } +// } +// +// input: +// set val(name), file(bam) from ch_bwa_bam +// +// output: +// set val(name), file("*.sorted.{bam,bam.bai}") into ch_sort_bam_merge +// file "*.{flagstat,idxstats,stats}" into ch_sort_bam_flagstat_mqc +// +// script: +// prefix = "${name}.Lb" +// """ +// samtools sort -@ $task.cpus -o ${prefix}.sorted.bam -T $name $bam +// samtools index ${prefix}.sorted.bam +// samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat +// samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats +// samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats +// """ +// } +// +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// /* -- -- */ +// /* -- MERGE LIBRARY BAM -- */ +// /* -- -- */ +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// +// /* +// * STEP 4.1 Merge BAM files for all libraries from same replicate +// */ +// ch_sort_bam_merge +// .map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] } +// .groupTuple(by: [0]) +// .map { it -> [ it[0], it[1].flatten() ] } +// .set { ch_sort_bam_merge } +// +// process MergedLibBAM { +// tag "$name" +// label 'process_medium' +// publishDir "${params.outdir}/bwa/mergedLibrary", mode: 'copy', +// saveAs: { filename -> +// if (filename.endsWith(".flagstat")) "samtools_stats/$filename" +// else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" +// else if (filename.endsWith(".stats")) "samtools_stats/$filename" +// else if (filename.endsWith(".metrics.txt")) "picard_metrics/$filename" +// else params.save_align_intermeds ? filename : null +// } +// +// input: +// set val(name), file(bams) from ch_sort_bam_merge +// +// output: +// set val(name), file("*${prefix}.sorted.{bam,bam.bai}") into ch_mlib_bam_filter, +// ch_mlib_bam_preseq, +// ch_mlib_bam_ataqv +// file "*.{flagstat,idxstats,stats}" into ch_mlib_bam_stats_mqc +// file "*.txt" into ch_mlib_bam_metrics_mqc +// +// script: +// prefix = "${name}.mLb.mkD" +// bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() +// def avail_mem = 3 +// if (!task.memory) { +// log.info "[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." +// } else { +// avail_mem = task.memory.toGiga() +// } +// if (bam_files.size() > 1) { +// """ +// picard -Xmx${avail_mem}g MergeSamFiles \\ +// ${'INPUT='+bam_files.join(' INPUT=')} \\ +// OUTPUT=${name}.sorted.bam \\ +// SORT_ORDER=coordinate \\ +// VALIDATION_STRINGENCY=LENIENT \\ +// TMP_DIR=tmp +// samtools index ${name}.sorted.bam +// +// picard -Xmx${avail_mem}g MarkDuplicates \\ +// INPUT=${name}.sorted.bam \\ +// OUTPUT=${prefix}.sorted.bam \\ +// ASSUME_SORTED=true \\ +// REMOVE_DUPLICATES=false \\ +// METRICS_FILE=${prefix}.MarkDuplicates.metrics.txt \\ +// VALIDATION_STRINGENCY=LENIENT \\ +// TMP_DIR=tmp +// +// samtools index ${prefix}.sorted.bam +// samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats +// samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat +// samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats +// """ +// } else { +// """ +// picard -Xmx${avail_mem}g MarkDuplicates \\ +// INPUT=${bam_files[0]} \\ +// OUTPUT=${prefix}.sorted.bam \\ +// ASSUME_SORTED=true \\ +// REMOVE_DUPLICATES=false \\ +// METRICS_FILE=${prefix}.MarkDuplicates.metrics.txt \\ +// VALIDATION_STRINGENCY=LENIENT \\ +// TMP_DIR=tmp +// +// samtools index ${prefix}.sorted.bam +// samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats +// samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat +// samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats +// """ +// } +// } +// +// /* +// * STEP 4.2 Filter BAM file at merged library-level +// */ +// process MergedLibBAMFilter { +// tag "$name" +// label 'process_medium' +// publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', +// saveAs: { filename -> +// if (params.single_end || params.save_align_intermeds) { +// if (filename.endsWith(".flagstat")) "samtools_stats/$filename" +// else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" +// else if (filename.endsWith(".stats")) "samtools_stats/$filename" +// else if (filename.endsWith(".sorted.bam")) filename +// else if (filename.endsWith(".sorted.bam.bai")) filename +// else null +// } +// } +// +// input: +// set val(name), file(bam) from ch_mlib_bam_filter +// file bed from ch_genome_filter_regions.collect() +// file bamtools_filter_config from ch_bamtools_filter_config +// +// output: +// set val(name), file("*.{bam,bam.bai}") into ch_mlib_filter_bam +// set val(name), file("*.flagstat") into ch_mlib_filter_bam_flagstat +// file "*.{idxstats,stats}" into ch_mlib_filter_bam_stats_mqc +// +// script: +// prefix = params.single_end ? "${name}.mLb.clN" : "${name}.mLb.flT" +// filter_params = params.single_end ? "-F 0x004" : "-F 0x004 -F 0x0008 -f 0x001" +// dup_params = params.keep_dups ? "" : "-F 0x0400" +// multimap_params = params.keep_multi_map ? "" : "-q 1" +// blacklist_params = params.blacklist ? "-L $bed" : "" +// name_sort_bam = params.single_end ? "" : "samtools sort -n -@ $task.cpus -o ${prefix}.bam -T $prefix ${prefix}.sorted.bam" +// """ +// samtools view \\ +// $filter_params \\ +// $dup_params \\ +// $multimap_params \\ +// $blacklist_params \\ +// -b ${bam[0]} \\ +// | bamtools filter \\ +// -out ${prefix}.sorted.bam \\ +// -script $bamtools_filter_config +// +// samtools index ${prefix}.sorted.bam +// samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat +// samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats +// samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats +// +// $name_sort_bam +// """ +// } +// +// /* +// * STEP 4.3 Remove orphan reads from paired-end BAM file +// */ +// if (params.single_end) { +// ch_mlib_filter_bam +// .into { ch_mlib_rm_orphan_bam_metrics; +// ch_mlib_rm_orphan_bam_bigwig; +// ch_mlib_rm_orphan_bam_macs; +// ch_mlib_rm_orphan_bam_plotfingerprint; +// ch_mlib_rm_orphan_bam_mrep; +// ch_mlib_name_bam_mlib_counts; +// ch_mlib_name_bam_mrep_counts } +// +// ch_mlib_filter_bam_flagstat +// .into { ch_mlib_rm_orphan_flagstat_bigwig; +// ch_mlib_rm_orphan_flagstat_macs; +// ch_mlib_rm_orphan_flagstat_mqc } +// +// ch_mlib_filter_bam_stats_mqc +// .set { ch_mlib_rm_orphan_stats_mqc } +// } else { +// process MergedLibBAMRemoveOrphan { +// tag "$name" +// label 'process_medium' +// publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', +// saveAs: { filename -> +// if (filename.endsWith(".flagstat")) "samtools_stats/$filename" +// else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" +// else if (filename.endsWith(".stats")) "samtools_stats/$filename" +// else if (filename.endsWith(".sorted.bam")) filename +// else if (filename.endsWith(".sorted.bam.bai")) filename +// else null +// } +// +// input: +// set val(name), file(bam) from ch_mlib_filter_bam +// +// output: +// set val(name), file("*.sorted.{bam,bam.bai}") into ch_mlib_rm_orphan_bam_metrics, +// ch_mlib_rm_orphan_bam_bigwig, +// ch_mlib_rm_orphan_bam_macs, +// ch_mlib_rm_orphan_bam_plotfingerprint, +// ch_mlib_rm_orphan_bam_mrep +// set val(name), file("${prefix}.bam") into ch_mlib_name_bam_mlib_counts, +// ch_mlib_name_bam_mrep_counts +// set val(name), file("*.flagstat") into ch_mlib_rm_orphan_flagstat_bigwig, +// ch_mlib_rm_orphan_flagstat_macs, +// ch_mlib_rm_orphan_flagstat_mqc +// file "*.{idxstats,stats}" into ch_mlib_rm_orphan_stats_mqc +// +// script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ +// prefix = "${name}.mLb.clN" +// """ +// bampe_rm_orphan.py ${bam[0]} ${prefix}.bam --only_fr_pairs +// +// samtools sort -@ $task.cpus -o ${prefix}.sorted.bam -T $prefix ${prefix}.bam +// samtools index ${prefix}.sorted.bam +// samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat +// samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats +// samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats +// """ +// } +// } +// +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// /* -- -- */ +// /* -- MERGE LIBRARY BAM POST-ANALYSIS -- */ +// /* -- -- */ +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// +// /* +// * STEP 5.1 preseq analysis after merging libraries and before filtering +// */ +// process MergedLibPreseq { +// tag "$name" +// label 'process_low' +// publishDir "${params.outdir}/bwa/mergedLibrary/preseq", mode: 'copy' +// +// when: +// !params.skip_preseq +// +// input: +// set val(name), file(bam) from ch_mlib_bam_preseq +// +// output: +// file "*.ccurve.txt" into ch_mlib_preseq_mqc +// +// script: +// prefix = "${name}.mLb.mkD" +// """ +// preseq lc_extrap -v -output ${prefix}.ccurve.txt -bam ${bam[0]} +// """ +// } +// +// /* +// * STEP 5.2 Picard CollectMultipleMetrics after merging libraries and filtering +// */ +// process MergedLibMetrics { +// tag "$name" +// label 'process_medium' +// publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', +// saveAs: { filename -> +// if (filename.endsWith("_metrics")) "picard_metrics/$filename" +// else if (filename.endsWith(".pdf")) "picard_metrics/pdf/$filename" +// else null +// } +// +// when: +// !params.skip_picard_metrics +// +// input: +// set val(name), file(bam) from ch_mlib_rm_orphan_bam_metrics +// file fasta from ch_fasta +// +// output: +// file "*_metrics" into ch_mlib_collectmetrics_mqc +// file "*.pdf" into ch_mlib_collectmetrics_pdf +// +// script: +// prefix = "${name}.mLb.clN" +// def avail_mem = 3 +// if (!task.memory) { +// log.info "[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." +// } else { +// avail_mem = task.memory.toGiga() +// } +// """ +// picard -Xmx${avail_mem}g CollectMultipleMetrics \\ +// INPUT=${bam[0]} \\ +// OUTPUT=${prefix}.CollectMultipleMetrics \\ +// REFERENCE_SEQUENCE=$fasta \\ +// VALIDATION_STRINGENCY=LENIENT \\ +// TMP_DIR=tmp +// """ +// } +// +// /* +// * STEP 5.3 Read depth normalised bigWig +// */ +// process MergedLibBigWig { +// tag "$name" +// label 'process_medium' +// publishDir "${params.outdir}/bwa/mergedLibrary/bigwig", mode: 'copy', +// saveAs: { filename -> +// if (filename.endsWith("scale_factor.txt")) "scale/$filename" +// else if (filename.endsWith(".bigWig")) "$filename" +// else null +// } +// +// input: +// set val(name), file(bam), file(flagstat) from ch_mlib_rm_orphan_bam_bigwig.join(ch_mlib_rm_orphan_flagstat_bigwig, by: [0]) +// file sizes from ch_genome_sizes_mlib_bigwig.collect() +// +// output: +// set val(name), file("*.bigWig") into ch_mlib_bigwig_plotprofile +// file "*scale_factor.txt" into ch_mlib_bigwig_scale +// file "*igv.txt" into ch_mlib_bigwig_igv +// +// script: +// prefix = "${name}.mLb.clN" +// pe_fragment = params.single_end ? "" : "-pc" +// extend = (params.single_end && params.fragment_size > 0) ? "-fs ${params.fragment_size}" : '' +// """ +// SCALE_FACTOR=\$(grep 'mapped (' $flagstat | awk '{print 1000000/\$1}') +// echo \$SCALE_FACTOR > ${prefix}.scale_factor.txt +// genomeCoverageBed -ibam ${bam[0]} -bg -scale \$SCALE_FACTOR $pe_fragment $extend | sort -T '.' -k1,1 -k2,2n > ${prefix}.bedGraph +// +// bedGraphToBigWig ${prefix}.bedGraph $sizes ${prefix}.bigWig +// +// find * -type f -name "*.bigWig" -exec echo -e "bwa/mergedLibrary/bigwig/"{}"\\t0,0,178" \\; > ${prefix}.bigWig.igv.txt +// """ +// } +// +// /* +// * STEP 5.4 generate gene body coverage plot with deepTools +// */ +// process MergedLibPlotProfile { +// tag "$name" +// label 'process_high' +// publishDir "${params.outdir}/bwa/mergedLibrary/deepTools/plotProfile", mode: 'copy' +// +// when: +// !params.skip_plot_profile +// +// input: +// set val(name), file(bigwig) from ch_mlib_bigwig_plotprofile +// file bed from ch_gene_bed +// +// output: +// file '*.{gz,pdf}' into ch_mlib_plotprofile_results +// file '*.plotProfile.tab' into ch_mlib_plotprofile_mqc +// +// script: +// prefix = "${name}.mLb.clN" +// """ +// computeMatrix scale-regions \\ +// --regionsFileName $bed \\ +// --scoreFileName $bigwig \\ +// --outFileName ${prefix}.computeMatrix.mat.gz \\ +// --outFileNameMatrix ${prefix}.computeMatrix.vals.mat.gz \\ +// --regionBodyLength 1000 \\ +// --beforeRegionStartLength 3000 \\ +// --afterRegionStartLength 3000 \\ +// --skipZeros \\ +// --smartLabels \\ +// --numberOfProcessors $task.cpus +// +// plotProfile --matrixFile ${prefix}.computeMatrix.mat.gz \\ +// --outFileName ${prefix}.plotProfile.pdf \\ +// --outFileNameData ${prefix}.plotProfile.tab +// """ +// } +// +// /* +// * STEP 5.5 deepTools plotFingerprint +// */ +// process MergedLibPlotFingerprint { +// tag "$name" +// label 'process_high' +// publishDir "${params.outdir}/bwa/mergedLibrary/deepTools/plotFingerprint", mode: 'copy' +// +// when: +// !params.skip_plot_fingerprint +// +// input: +// set val(name), file(bam) from ch_mlib_rm_orphan_bam_plotfingerprint +// +// output: +// file '*.{txt,pdf}' into ch_mlib_plotfingerprint_results +// file '*.raw.txt' into ch_mlib_plotfingerprint_mqc +// +// script: +// prefix = "${name}.mLb.clN" +// extend = (params.single_end && params.fragment_size > 0) ? "--extendReads ${params.fragment_size}" : '' +// """ +// plotFingerprint \\ +// --bamfiles ${bam[0]} \\ +// --plotFile ${prefix}.plotFingerprint.pdf \\ +// $extend \\ +// --labels $prefix \\ +// --outRawCounts ${prefix}.plotFingerprint.raw.txt \\ +// --outQualityMetrics ${prefix}.plotFingerprint.qcmetrics.txt \\ +// --skipZeros \\ +// --numberOfProcessors $task.cpus \\ +// --numberOfSamples $params.fingerprint_bins +// """ +// } +// +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// /* -- -- */ +// /* -- MERGE LIBRARY PEAK ANALYSIS -- */ +// /* -- -- */ +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// +// /* +// * STEP 6.1 Call peaks with MACS2 and calculate FRiP score +// */ +// process MergedLibMACSCallPeak { +// tag "$name" +// label 'process_medium' +// publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}", mode: 'copy', +// saveAs: { filename -> +// if (filename.endsWith(".tsv")) "qc/$filename" +// else if (filename.endsWith(".igv.txt")) null +// else filename +// } +// +// when: +// params.macs_gsize +// +// input: +// set val(name), file(bam), file(flagstat) from ch_mlib_rm_orphan_bam_macs.join(ch_mlib_rm_orphan_flagstat_macs, by: [0]) +// file mlib_peak_count_header from ch_mlib_peak_count_header +// file mlib_frip_score_header from ch_mlib_frip_score_header +// +// output: +// set val(name), file("*.{bed,xls,gappedPeak,bdg}") into ch_mlib_macs_output +// set val(name), file("*$PEAK_TYPE") into ch_mlib_macs_homer, +// ch_mlib_macs_qc, +// ch_mlib_macs_consensus, +// ch_mlib_macs_ataqv +// file "*igv.txt" into ch_mlib_macs_igv +// file "*_mqc.tsv" into ch_mlib_macs_mqc +// +// script: +// prefix = "${name}.mLb.clN" +// broad = params.narrow_peak ? '' : "--broad --broad-cutoff ${params.broad_cutoff}" +// format = params.single_end ? "BAM" : "BAMPE" +// pileup = params.save_macs_pileup ? "-B --SPMR" : "" +// """ +// macs2 callpeak \\ +// -t ${bam[0]} \\ +// $broad \\ +// -f $format \\ +// -g $params.macs_gsize \\ +// -n $prefix \\ +// $pileup \\ +// --keep-dup all \\ +// --nomodel +// +// cat ${prefix}_peaks.${PEAK_TYPE} | wc -l | awk -v OFS='\t' '{ print "${name}", \$1 }' | cat $mlib_peak_count_header - > ${prefix}_peaks.count_mqc.tsv +// +// READS_IN_PEAKS=\$(intersectBed -a ${bam[0]} -b ${prefix}_peaks.${PEAK_TYPE} -bed -c -f 0.20 | awk -F '\t' '{sum += \$NF} END {print sum}') +// grep 'mapped (' $flagstat | awk -v a="\$READS_IN_PEAKS" -v OFS='\t' '{print "${name}", a/\$1}' | cat $mlib_frip_score_header - > ${prefix}_peaks.FRiP_mqc.tsv +// +// find * -type f -name "*.${PEAK_TYPE}" -exec echo -e "bwa/mergedLibrary/macs/${PEAK_TYPE}/"{}"\\t0,0,178" \\; > ${prefix}_peaks.igv.txt +// """ +// } +// +// /* +// * STEP 6.2 Annotate peaks with HOMER +// */ +// process MergedLibAnnotatePeaks { +// tag "$name" +// label "process_medium" +// publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}", mode: 'copy' +// +// when: +// params.macs_gsize +// +// input: +// set val(name), file(peak) from ch_mlib_macs_homer +// file fasta from ch_fasta +// file gtf from ch_gtf +// +// output: +// file "*.txt" into ch_mlib_macs_annotate +// +// script: +// prefix = "${name}.mLb.clN" +// """ +// annotatePeaks.pl \\ +// $peak \\ +// $fasta \\ +// -gid \\ +// -gtf $gtf \\ +// -cpu $task.cpus \\ +// > ${prefix}_peaks.annotatePeaks.txt +// """ +// } +// +// /* +// * STEP 6.3 Aggregated QC plots for peaks, FRiP and peak-to-gene annotation +// */ +// process MergedLibPeakQC { +// label "process_medium" +// publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/qc", mode: 'copy' +// +// when: +// params.macs_gsize +// +// input: +// file peaks from ch_mlib_macs_qc.collect{ it[1] } +// file annos from ch_mlib_macs_annotate.collect() +// file mlib_peak_annotation_header from ch_mlib_peak_annotation_header +// +// output: +// file "*.{txt,pdf}" into ch_mlib_peak_qc +// file "*.tsv" into ch_mlib_peak_qc_mqc +// +// script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ +// suffix = 'mLb.clN' +// """ +// plot_macs_qc.r \\ +// -i ${peaks.join(',')} \\ +// -s ${peaks.join(',').replaceAll(".${suffix}_peaks.${PEAK_TYPE}","")} \\ +// -o ./ \\ +// -p macs_peak.${suffix} +// +// plot_homer_annotatepeaks.r \\ +// -i ${annos.join(',')} \\ +// -s ${annos.join(',').replaceAll(".${suffix}_peaks.annotatePeaks.txt","")} \\ +// -o ./ \\ +// -p macs_annotatePeaks.${suffix} +// +// cat $mlib_peak_annotation_header macs_annotatePeaks.${suffix}.summary.txt > macs_annotatePeaks.${suffix}.summary_mqc.tsv +// """ +// } +// +// /* +// * STEP 6.4 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection +// */ +// process MergedLibConsensusPeakSet { +// label 'process_long' +// publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: 'copy', +// saveAs: { filename -> +// if (filename.endsWith(".igv.txt")) null +// else filename +// } +// +// when: +// params.macs_gsize && (replicatesExist || multipleGroups) && !params.skip_consensus_peaks +// +// input: +// file peaks from ch_mlib_macs_consensus.collect{ it[1] } +// +// output: +// file "*.bed" into ch_mlib_macs_consensus_bed +// file "*.saf" into ch_mlib_macs_consensus_saf +// file "*.boolean.txt" into ch_mlib_macs_consensus_bool +// file "*.intersect.{txt,plot.pdf}" into ch_mlib_macs_consensus_intersect +// file "*igv.txt" into ch_mlib_macs_consensus_igv +// +// script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ +// suffix = 'mLb.clN' +// prefix = "consensus_peaks.${suffix}" +// mergecols = params.narrow_peak ? (2..10).join(',') : (2..9).join(',') +// collapsecols = params.narrow_peak ? (["collapse"]*9).join(',') : (["collapse"]*8).join(',') +// expandparam = params.narrow_peak ? "--is_narrow_peak" : "" +// """ +// sort -T '.' -k1,1 -k2,2n ${peaks.collect{it.toString()}.sort().join(' ')} \\ +// | mergeBed -c $mergecols -o $collapsecols > ${prefix}.txt +// +// macs2_merged_expand.py \\ +// ${prefix}.txt \\ +// ${peaks.collect{it.toString()}.sort().join(',').replaceAll("_peaks.${PEAK_TYPE}","")} \\ +// ${prefix}.boolean.txt \\ +// --min_replicates $params.min_reps_consensus \\ +// $expandparam +// +// awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$1, \$2, \$3, \$4, "0", "+" }' ${prefix}.boolean.txt > ${prefix}.bed +// +// echo -e "GeneID\tChr\tStart\tEnd\tStrand" > ${prefix}.saf +// awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$4, \$1, \$2, \$3, "+" }' ${prefix}.boolean.txt >> ${prefix}.saf +// +// sed -i 's/.${suffix}//g' ${prefix}.boolean.intersect.txt +// plot_peak_intersect.r -i ${prefix}.boolean.intersect.txt -o ${prefix}.boolean.intersect.plot.pdf +// +// find * -type f -name "${prefix}.bed" -exec echo -e "bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus/"{}"\\t0,0,0" \\; > ${prefix}.bed.igv.txt +// """ +// } +// +// /* +// * STEP 6.5 Annotate consensus peaks with HOMER, and add annotation to boolean output file +// */ +// process MergedLibConsensusPeakSetAnnotate { +// label "process_medium" +// publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: 'copy' +// +// when: +// params.macs_gsize && (replicatesExist || multipleGroups) && !params.skip_consensus_peaks +// +// input: +// file bed from ch_mlib_macs_consensus_bed +// file bool from ch_mlib_macs_consensus_bool +// file fasta from ch_fasta +// file gtf from ch_gtf +// +// output: +// file "*.annotatePeaks.txt" into ch_mlib_macs_consensus_annotate +// +// script: +// prefix = "consensus_peaks.mLb.clN" +// """ +// annotatePeaks.pl \\ +// $bed \\ +// $fasta \\ +// -gid \\ +// -gtf $gtf \\ +// -cpu $task.cpus \\ +// > ${prefix}.annotatePeaks.txt +// +// cut -f2- ${prefix}.annotatePeaks.txt | awk 'NR==1; NR > 1 {print \$0 | "sort -T '.' -k1,1 -k2,2n"}' | cut -f6- > tmp.txt +// paste $bool tmp.txt > ${prefix}.boolean.annotatePeaks.txt +// """ +// } +// +// /* +// * STEP 6.6 Count reads in consensus peaks with featureCounts +// */ +// process MergedLibConsensusPeakSetCounts { +// label 'process_medium' +// publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: 'copy' +// +// when: +// params.macs_gsize && (replicatesExist || multipleGroups) && !params.skip_consensus_peaks +// +// input: +// file bams from ch_mlib_name_bam_mlib_counts.collect{ it[1] } +// file saf from ch_mlib_macs_consensus_saf.collect() +// +// output: +// file "*featureCounts.txt" into ch_mlib_macs_consensus_counts +// file "*featureCounts.txt.summary" into ch_mlib_macs_consensus_counts_mqc +// +// script: +// prefix = "consensus_peaks.mLb.clN" +// bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() +// pe_params = params.single_end ? '' : "-p --donotsort" +// """ +// featureCounts \\ +// -F SAF \\ +// -O \\ +// --fracOverlap 0.2 \\ +// -T $task.cpus \\ +// $pe_params \\ +// -a $saf \\ +// -o ${prefix}.featureCounts.txt \\ +// ${bam_files.join(' ')} +// """ +// } +// +// /* +// * STEP 6.7 Perform differential analysis with DESeq2 +// */ +// process MergedLibConsensusPeakSetDESeq { +// label 'process_medium' +// publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus/deseq2", mode: 'copy', +// saveAs: { filename -> +// if (filename.endsWith(".igv.txt")) null +// else filename +// } +// +// when: +// params.macs_gsize && replicatesExist && multipleGroups && !params.skip_consensus_peaks && !params.skip_diff_analysis +// +// input: +// file counts from ch_mlib_macs_consensus_counts +// file mlib_deseq2_pca_header from ch_mlib_deseq2_pca_header +// file mlib_deseq2_clustering_header from ch_mlib_deseq2_clustering_header +// +// output: +// file "*.{RData,results.txt,pdf,log}" into ch_mlib_macs_consensus_deseq_results +// file "sizeFactors" into ch_mlib_macs_consensus_deseq_factors +// file "*vs*/*.{pdf,txt}" into ch_mlib_macs_consensus_deseq_comp_results +// file "*vs*/*.bed" into ch_mlib_macs_consensus_deseq_comp_bed +// file "*igv.txt" into ch_mlib_macs_consensus_deseq_comp_igv +// file "*.tsv" into ch_mlib_macs_consensus_deseq_mqc +// +// script: +// prefix = "consensus_peaks.mLb.clN" +// bam_ext = params.single_end ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" +// """ +// featurecounts_deseq2.r \\ +// --featurecount_file $counts \\ +// --bam_suffix '$bam_ext' \\ +// --outdir ./ \\ +// --outprefix $prefix \\ +// --outsuffix .mLb \\ +// --cores $task.cpus +// +// cat $mlib_deseq2_pca_header ${prefix}.pca.vals.txt > ${prefix}.pca.vals_mqc.tsv +// cat $mlib_deseq2_clustering_header ${prefix}.sample.dists.txt > ${prefix}.sample.dists_mqc.tsv +// +// find * -type f -name "*.FDR0.05.results.bed" -exec echo -e "bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus/deseq2/"{}"\\t255,0,0" \\; > ${prefix}.igv.txt +// """ +// } +// +// /* +// * STEP 6.8 Run ataqv on BAM file and corresponding peaks +// */ +// process MergedLibAtaqv { +// tag "$name" +// label 'process_medium' +// publishDir "${params.outdir}/bwa/mergedLibrary/ataqv/${PEAK_TYPE}", mode: 'copy' +// +// when: +// !params.skip_ataqv +// +// input: +// set val(name), file(bam), file(peak) from ch_mlib_bam_ataqv.join(ch_mlib_macs_ataqv, by: [0]) +// file autosomes from ch_genome_autosomes.collect() +// file tss_bed from ch_tss_bed +// +// output: +// file "*.json" into ch_mlib_ataqv +// +// script: +// peak_param = params.macs_gsize ? "--peak-file ${peak}" : "" +// mito_param = params.mito_name ? "--mitochondrial-reference-name ${params.mito_name}" : "" +// """ +// ataqv \\ +// --threads $task.cpus \\ +// $peak_param \\ +// --tss-file $tss_bed \\ +// --metrics-file ${name}.ataqv.json \\ +// --name $name \\ +// --ignore-read-groups \\ +// --autosomal-reference-file $autosomes \\ +// $mito_param \\ +// NA \\ +// ${bam[0]} +// """ +// } +// +// /* +// * STEP 6.9 run ataqv mkarv on all JSON files to render web app +// */ +// process MergedLibAtaqvMkarv { +// label 'process_medium' +// publishDir "${params.outdir}/bwa/mergedLibrary/ataqv/${PEAK_TYPE}", mode: 'copy' +// +// when: +// !params.skip_ataqv +// +// input: +// file json from ch_mlib_ataqv.collect() +// +// output: +// file "html" into ch_mlib_ataqv_mkarv +// +// script: +// """ +// mkarv \\ +// --concurrency $task.cpus \\ +// --force \\ +// ./html/ \\ +// ${json.join(' ')} +// """ +// } +// +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// /* -- -- */ +// /* -- MERGE REPLICATE BAM -- */ +// /* -- -- */ +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// +// /* +// * STEP 7 Merge library BAM files across all replicates +// */ +// ch_mlib_rm_orphan_bam_mrep +// .map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] } +// .groupTuple(by: [0]) +// .map { it -> [ it[0], it[1].flatten() ] } +// .set { ch_mlib_rm_orphan_bam_mrep } +// +// process MergedRepBAM { +// tag "$name" +// label 'process_medium' +// publishDir "${params.outdir}/bwa/mergedReplicate", mode: 'copy', +// saveAs: { filename -> +// if (filename.endsWith(".flagstat")) "samtools_stats/$filename" +// else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" +// else if (filename.endsWith(".stats")) "samtools_stats/$filename" +// else if (filename.endsWith(".metrics.txt")) "picard_metrics/$filename" +// else filename +// } +// +// input: +// set val(name), file(bams) from ch_mlib_rm_orphan_bam_mrep +// +// output: +// set val(name), file("*${prefix}.sorted.{bam,bam.bai}") into ch_mrep_bam_bigwig, +// ch_mrep_bam_macs +// set val(name), file("*.flagstat") into ch_mrep_bam_flagstat_bigwig, +// ch_mrep_bam_flagstat_macs, +// ch_mrep_bam_flagstat_mqc +// file "*.{idxstats,stats}" into ch_mrep_bam_stats_mqc +// file "*.txt" into ch_mrep_bam_metrics_mqc +// +// when: +// !params.skip_merge_replicates && replicatesExist +// +// script: +// prefix = "${name}.mRp.clN" +// bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() +// def avail_mem = 3 +// if (!task.memory) { +// log.info "[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." +// } else { +// avail_mem = task.memory.toGiga() +// } +// if (bam_files.size() > 1) { +// """ +// picard -Xmx${avail_mem}g MergeSamFiles \\ +// ${'INPUT='+bam_files.join(' INPUT=')} \\ +// OUTPUT=${name}.sorted.bam \\ +// SORT_ORDER=coordinate \\ +// VALIDATION_STRINGENCY=LENIENT \\ +// TMP_DIR=tmp +// samtools index ${name}.sorted.bam +// +// picard -Xmx${avail_mem}g MarkDuplicates \\ +// INPUT=${name}.sorted.bam \\ +// OUTPUT=${prefix}.sorted.bam \\ +// ASSUME_SORTED=true \\ +// REMOVE_DUPLICATES=true \\ +// METRICS_FILE=${prefix}.MarkDuplicates.metrics.txt \\ +// VALIDATION_STRINGENCY=LENIENT \\ +// TMP_DIR=tmp +// +// samtools index ${prefix}.sorted.bam +// samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat +// samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats +// samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats +// """ +// } else { +// """ +// ln -s ${bams[0]} ${prefix}.sorted.bam +// ln -s ${bams[1]} ${prefix}.sorted.bam.bai +// touch ${prefix}.MarkDuplicates.metrics.txt +// samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat +// samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats +// samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats +// """ +// } +// } +// +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// /* -- -- */ +// /* -- MERGE REPLICATE BAM POST-ANALYSIS -- */ +// /* -- -- */ +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// +// /* +// * STEP 8.1 Read depth normalised bigWig +// */ +// process MergedRepBigWig { +// tag "$name" +// label 'process_medium' +// publishDir "${params.outdir}/bwa/mergedReplicate/bigwig", mode: 'copy', +// saveAs: { filename -> +// if (filename.endsWith("scale_factor.txt")) "scale/$filename" +// else if (filename.endsWith(".bigWig")) "$filename" +// else null +// } +// +// when: +// !params.skip_merge_replicates && replicatesExist +// +// input: +// set val(name), file(bam), file(flagstat) from ch_mrep_bam_bigwig.join(ch_mrep_bam_flagstat_bigwig, by: [0]) +// file sizes from ch_genome_sizes_mrep_bigwig.collect() +// +// output: +// set val(name), file("*.bigWig") into ch_mrep_bigwig +// file "*scale_factor.txt" into ch_mrep_bigwig_scale +// file "*igv.txt" into ch_mrep_bigwig_igv +// +// script: +// prefix = "${name}.mRp.clN" +// pe_fragment = params.single_end ? "" : "-pc" +// extend = (params.single_end && params.fragment_size > 0) ? "-fs ${params.fragment_size}" : '' +// """ +// SCALE_FACTOR=\$(grep 'mapped (' $flagstat | awk '{print 1000000/\$1}') +// echo \$SCALE_FACTOR > ${prefix}.scale_factor.txt +// genomeCoverageBed -ibam ${bam[0]} -bg -scale \$SCALE_FACTOR $pe_fragment $extend | sort -T '.' -k1,1 -k2,2n > ${prefix}.bedGraph +// +// bedGraphToBigWig ${prefix}.bedGraph $sizes ${prefix}.bigWig +// +// find * -type f -name "*.bigWig" -exec echo -e "bwa/mergedReplicate/bigwig/"{}"\\t0,0,178" \\; > ${prefix}.bigWig.igv.txt +// """ +// } +// +// /* +// * STEP 8.2 Call peaks with MACS2 and calculate FRiP score +// */ +// process MergedRepMACSCallPeak { +// tag "$name" +// label 'process_medium' +// publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}", mode: 'copy', +// saveAs: { filename -> +// if (filename.endsWith(".tsv")) "qc/$filename" +// else if (filename.endsWith(".igv.txt")) null +// else filename +// } +// +// when: +// !params.skip_merge_replicates && replicatesExist && params.macs_gsize +// +// input: +// set val(name), file(bam), file(flagstat) from ch_mrep_bam_macs.join(ch_mrep_bam_flagstat_macs, by: [0]) +// file mrep_peak_count_header from ch_mrep_peak_count_header +// file mrep_frip_score_header from ch_mrep_frip_score_header +// +// output: +// set val(name), file("*.{bed,xls,gappedPeak,bdg}") into ch_mrep_macs_output +// set val(name), file("*$PEAK_TYPE") into ch_mrep_macs_homer, +// ch_mrep_macs_qc, +// ch_mrep_macs_consensus +// file "*igv.txt" into ch_mrep_macs_igv +// file "*_mqc.tsv" into ch_mrep_macs_mqc +// +// script: +// prefix = "${name}.mRp.clN" +// broad = params.narrow_peak ? '' : "--broad --broad-cutoff ${params.broad_cutoff}" +// format = params.single_end ? "BAM" : "BAMPE" +// pileup = params.save_macs_pileup ? "-B --SPMR" : "" +// """ +// macs2 callpeak \\ +// -t ${bam[0]} \\ +// $broad \\ +// -f $format \\ +// -g $params.macs_gsize \\ +// -n $prefix \\ +// $pileup \\ +// --keep-dup all \\ +// --nomodel +// +// cat ${prefix}_peaks.${PEAK_TYPE} | wc -l | awk -v OFS='\t' '{ print "${name}", \$1 }' | cat $mrep_peak_count_header - > ${prefix}_peaks.count_mqc.tsv +// +// READS_IN_PEAKS=\$(intersectBed -a ${bam[0]} -b ${prefix}_peaks.${PEAK_TYPE} -bed -c -f 0.20 | awk -F '\t' '{sum += \$NF} END {print sum}') +// grep 'mapped (' $flagstat | awk -v a="\$READS_IN_PEAKS" -v OFS='\t' '{print "${name}", a/\$1}' | cat $mrep_frip_score_header - > ${prefix}_peaks.FRiP_mqc.tsv +// +// find * -type f -name "*.${PEAK_TYPE}" -exec echo -e "bwa/mergedReplicate/macs/${PEAK_TYPE}/"{}"\\t0,0,178" \\; > ${prefix}_peaks.igv.txt +// """ +// } +// +// /* +// * STEP 8.3 Annotate peaks with HOMER +// */ +// process MergedRepAnnotatePeaks { +// tag "$name" +// label "process_medium" +// publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}", mode: 'copy' +// +// when: +// !params.skip_merge_replicates && replicatesExist && params.macs_gsize +// +// input: +// set val(name), file(peak) from ch_mrep_macs_homer +// file fasta from ch_fasta +// file gtf from ch_gtf +// +// output: +// file "*.txt" into ch_mrep_macs_annotate +// +// script: +// prefix = "${name}.mRp.clN" +// """ +// annotatePeaks.pl \\ +// $peak \\ +// $fasta \\ +// -gid \\ +// -gtf $gtf \\ +// -cpu $task.cpus \\ +// > ${prefix}_peaks.annotatePeaks.txt +// """ +// } +// +// /* +// * STEP 8.4 Aggregated QC plots for peaks, FRiP and peak-to-gene annotation +// */ +// process MergedRepPeakQC { +// label "process_medium" +// publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/qc", mode: 'copy' +// +// when: +// !params.skip_merge_replicates && replicatesExist && params.macs_gsize +// +// input: +// file peaks from ch_mrep_macs_qc.collect{ it[1] } +// file annos from ch_mrep_macs_annotate.collect() +// file mrep_peak_annotation_header from ch_mrep_peak_annotation_header +// +// output: +// file "*.{txt,pdf}" into ch_mrep_peak_qc +// file "*.tsv" into ch_mrep_peak_qc_mqc +// +// script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ +// suffix = 'mRp.clN' +// """ +// plot_macs_qc.r \\ +// -i ${peaks.join(',')} \\ +// -s ${peaks.join(',').replaceAll(".${suffix}_peaks.${PEAK_TYPE}","")} \\ +// -o ./ \\ +// -p macs_peak.${suffix} +// +// plot_homer_annotatepeaks.r \\ +// -i ${annos.join(',')} \\ +// -s ${annos.join(',').replaceAll(".${suffix}_peaks.annotatePeaks.txt","")} \\ +// -o ./ \\ +// -p macs_annotatePeaks.${suffix} +// +// cat $mrep_peak_annotation_header macs_annotatePeaks.${suffix}.summary.txt > macs_annotatePeaks.${suffix}.summary_mqc.tsv +// """ +// } +// +// /* +// * STEP 8.5 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection +// */ +// process MergedRepConsensusPeakSet { +// label 'process_long' +// publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: 'copy', +// saveAs: { filename -> +// if (filename.endsWith(".igv.txt")) null +// else filename +// } +// +// when: +// !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks +// +// input: +// file peaks from ch_mrep_macs_consensus.collect{ it[1] } +// +// output: +// file "*.bed" into ch_mrep_macs_consensus_bed +// file "*.saf" into ch_mrep_macs_consensus_saf +// file "*.boolean.txt" into ch_mrep_macs_consensus_bool +// file "*.intersect.{txt,plot.pdf}" into ch_mrep_macs_consensus_intersect +// file "*igv.txt" into ch_mrep_macs_consensus_igv +// +// script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ +// suffix = 'mRp.clN' +// prefix = "consensus_peaks.${suffix}" +// mergecols = params.narrow_peak ? (2..10).join(',') : (2..9).join(',') +// collapsecols = params.narrow_peak ? (["collapse"]*9).join(',') : (["collapse"]*8).join(',') +// expandparam = params.narrow_peak ? "--is_narrow_peak" : "" +// """ +// sort -T '.' -k1,1 -k2,2n ${peaks.collect{it.toString()}.sort().join(' ')} \\ +// | mergeBed -c $mergecols -o $collapsecols > ${prefix}.txt +// +// macs2_merged_expand.py \\ +// ${prefix}.txt \\ +// ${peaks.collect{it.toString()}.sort().join(',').replaceAll("_peaks.${PEAK_TYPE}","")} \\ +// ${prefix}.boolean.txt \\ +// $expandparam +// +// awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$1, \$2, \$3, \$4, "0", "+" }' ${prefix}.boolean.txt > ${prefix}.bed +// +// echo -e "GeneID\tChr\tStart\tEnd\tStrand" > ${prefix}.saf +// awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$4, \$1, \$2, \$3, "+" }' ${prefix}.boolean.txt >> ${prefix}.saf +// +// sed -i 's/.${suffix}//g' ${prefix}.boolean.intersect.txt +// plot_peak_intersect.r -i ${prefix}.boolean.intersect.txt -o ${prefix}.boolean.intersect.plot.pdf +// +// find * -type f -name "${prefix}.bed" -exec echo -e "bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus/"{}"\\t0,0,0" \\; > ${prefix}.bed.igv.txt +// """ +// } +// +// /* +// * STEP 8.6 Annotate consensus peaks with HOMER, and add annotation to boolean output file +// */ +// process MergedRepConsensusPeakSetAnnotate { +// label "process_medium" +// publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: 'copy' +// +// when: +// !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks +// +// input: +// file bed from ch_mrep_macs_consensus_bed +// file bool from ch_mrep_macs_consensus_bool +// file fasta from ch_fasta +// file gtf from ch_gtf +// +// output: +// file "*.annotatePeaks.txt" into ch_mrep_macs_consensus_annotate +// +// script: +// prefix = "consensus_peaks.mRp.clN" +// """ +// annotatePeaks.pl \\ +// $bed \\ +// $fasta \\ +// -gid \\ +// -gtf $gtf \\ +// -cpu $task.cpus \\ +// > ${prefix}.annotatePeaks.txt +// +// cut -f2- ${prefix}.annotatePeaks.txt | awk 'NR==1; NR > 1 {print \$0 | "sort -T '.' -k1,1 -k2,2n"}' | cut -f6- > tmp.txt +// paste $bool tmp.txt > ${prefix}.boolean.annotatePeaks.txt +// """ +// } +// +// /* +// * STEP 8.7 Count reads in consensus peaks with featureCounts +// */ +// process MergedRepConsensusPeakSetCounts { +// label 'process_medium' +// publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: 'copy' +// +// when: +// !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks +// +// input: +// file bams from ch_mlib_name_bam_mrep_counts.collect{ it[1] } +// file saf from ch_mrep_macs_consensus_saf.collect() +// +// output: +// file "*featureCounts.txt" into ch_mrep_macs_consensus_counts +// file "*featureCounts.txt.summary" into ch_mrep_macs_consensus_counts_mqc +// +// script: +// prefix = "consensus_peaks.mRp.clN" +// bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() +// pe_params = params.single_end ? '' : "-p --donotsort" +// """ +// featureCounts \\ +// -F SAF \\ +// -O \\ +// --fracOverlap 0.2 \\ +// -T $task.cpus \\ +// $pe_params \\ +// -a $saf \\ +// -o ${prefix}.featureCounts.txt \\ +// ${bam_files.join(' ')} +// """ +// } +// +// /* +// * STEP 8.8 Perform differential analysis with DESeq2 +// */ +// process MergedRepConsensusPeakSetDESeq { +// label 'process_medium' +// publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus/deseq2", mode: 'copy', +// saveAs: { filename -> +// if (filename.endsWith(".igv.txt")) null +// else filename +// } +// +// when: +// !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks && !params.skip_diff_analysis +// +// input: +// file counts from ch_mrep_macs_consensus_counts +// file mrep_deseq2_pca_header from ch_mrep_deseq2_pca_header +// file mrep_deseq2_clustering_header from ch_mrep_deseq2_clustering_header +// +// output: +// file "*.{RData,results.txt,pdf,log}" into ch_mrep_macs_consensus_deseq_results +// file "sizeFactors" into ch_mrep_macs_consensus_deseq_factors +// file "*vs*/*.{pdf,txt}" into ch_mrep_macs_consensus_deseq_comp_results +// file "*vs*/*.bed" into ch_mrep_macs_consensus_deseq_comp_bed +// file "*igv.txt" into ch_mrep_macs_consensus_deseq_comp_igv +// file "*.tsv" into ch_mrep_macs_consensus_deseq_mqc +// +// script: +// prefix = "consensus_peaks.mRp.clN" +// bam_ext = params.single_end ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" +// """ +// featurecounts_deseq2.r \\ +// --featurecount_file $counts \\ +// --bam_suffix '$bam_ext' \\ +// --outdir ./ \\ +// --outprefix $prefix \\ +// --outsuffix .mRp \\ +// --cores $task.cpus +// +// cat $mrep_deseq2_pca_header ${prefix}.pca.vals.txt > ${prefix}.pca.vals_mqc.tsv +// cat $mrep_deseq2_clustering_header ${prefix}.sample.dists.txt > ${prefix}.sample.dists_mqc.tsv +// +// find * -type f -name "*.FDR0.05.results.bed" -exec echo -e "bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus/deseq2/"{}"\\t255,0,0" \\; > ${prefix}.igv.txt +// """ +// } +// +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// /* -- -- */ +// /* -- IGV -- */ +// /* -- -- */ +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// +// /* +// * STEP 9 - Create IGV session file +// */ +// process IGV { +// publishDir "${params.outdir}/igv/${PEAK_TYPE}", mode: 'copy' +// +// when: +// !params.skip_igv +// +// input: +// file fasta from ch_fasta +// +// file bigwigs from ch_mlib_bigwig_igv.collect().ifEmpty([]) +// file peaks from ch_mlib_macs_igv.collect().ifEmpty([]) +// file consensus_peaks from ch_mlib_macs_consensus_igv.collect().ifEmpty([]) +// file differential_peaks from ch_mlib_macs_consensus_deseq_comp_igv.collect().ifEmpty([]) +// +// file rbigwigs from ch_mrep_bigwig_igv.collect().ifEmpty([]) +// file rpeaks from ch_mrep_macs_igv.collect().ifEmpty([]) +// file rconsensus_peaks from ch_mrep_macs_consensus_igv.collect().ifEmpty([]) +// file rdifferential_peaks from ch_mrep_macs_consensus_deseq_comp_igv.collect().ifEmpty([]) +// +// output: +// file "*.{txt,xml}" into ch_igv_session +// +// script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ +// """ +// cat *.txt > igv_files.txt +// igv_files_to_session.py igv_session.xml igv_files.txt ../../genome/${fasta.getName()} --path_prefix '../../' +// """ +// } +// +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// /* -- -- */ +// /* -- MULTIQC -- */ +// /* -- -- */ +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// +// /* +// * Parse software version numbers +// */ +// process get_software_versions { +// publishDir "${params.outdir}/pipeline_info", mode: 'copy', +// saveAs: { filename -> +// if (filename.indexOf(".csv") > 0) filename +// else null +// } +// +// output: +// file 'software_versions_mqc.yaml' into ch_software_versions_mqc +// file "software_versions.csv" +// +// script: +// """ +// echo $workflow.manifest.version > v_pipeline.txt +// echo $workflow.nextflow.version > v_nextflow.txt +// fastqc --version > v_fastqc.txt +// trim_galore --version > v_trim_galore.txt +// echo \$(bwa 2>&1) > v_bwa.txt +// samtools --version > v_samtools.txt +// bedtools --version > v_bedtools.txt +// echo \$(bamtools --version 2>&1) > v_bamtools.txt +// echo \$(plotFingerprint --version 2>&1) > v_deeptools.txt || true +// picard MarkDuplicates --version &> v_picard.txt || true +// echo \$(R --version 2>&1) > v_R.txt +// python -c "import pysam; print(pysam.__version__)" > v_pysam.txt +// echo \$(macs2 --version 2>&1) > v_macs2.txt +// touch v_homer.txt +// echo \$(ataqv --version 2>&1) > v_ataqv.txt +// echo \$(featureCounts -v 2>&1) > v_featurecounts.txt +// preseq &> v_preseq.txt +// multiqc --version > v_multiqc.txt +// scrape_software_versions.py &> software_versions_mqc.yaml +// """ +// } +// +// Channel.from(summary.collect{ [it.key, it.value] }) +// .map { k,v -> "
$k
${v ?: 'N/A'}
" } +// .reduce { a, b -> return [a, b].join("\n ") } +// .map { x -> """ +// id: 'nf-core-chipseq-summary' +// description: " - this information is collected when the pipeline is started." +// section_name: 'nf-core/chipseq Workflow Summary' +// section_href: 'https://github.com/nf-core/chipseq' +// plot_type: 'html' +// data: | +//
+// $x +//
+// """.stripIndent() } +// .set { ch_workflow_summary } +// +// /* +// * STEP 10 - MultiQC +// */ +// process MultiQC { +// publishDir "${params.outdir}/multiqc/${PEAK_TYPE}", mode: 'copy' +// +// when: +// !params.skip_multiqc +// +// input: +// file (multiqc_config) from ch_multiqc_config +// file (mqc_custom_config) from ch_multiqc_custom_config.collect().ifEmpty([]) +// +// file ('software_versions/*') from ch_software_versions_mqc.collect() +// file workflow_summary from ch_workflow_summary.collectFile(name: "workflow_summary_mqc.yaml") +// +// file ('fastqc/*') from ch_fastqc_reports_mqc.collect().ifEmpty([]) +// file ('trimgalore/*') from ch_trimgalore_results_mqc.collect().ifEmpty([]) +// file ('trimgalore/fastqc/*') from ch_trimgalore_fastqc_reports_mqc.collect().ifEmpty([]) +// +// file ('alignment/library/*') from ch_sort_bam_flagstat_mqc.collect() +// +// file ('alignment/mergedLibrary/*') from ch_mlib_bam_stats_mqc.collect() +// file ('alignment/mergedLibrary/*') from ch_mlib_rm_orphan_flagstat_mqc.collect{it[1]} +// file ('alignment/mergedLibrary/*') from ch_mlib_rm_orphan_stats_mqc.collect() +// file ('alignment/mergedLibrary/picard_metrics/*') from ch_mlib_bam_metrics_mqc.collect() +// file ('alignment/mergedLibrary/picard_metrics/*') from ch_mlib_collectmetrics_mqc.collect() +// file ('macs/mergedLibrary/*') from ch_mlib_macs_mqc.collect().ifEmpty([]) +// file ('macs/mergedLibrary/*') from ch_mlib_peak_qc_mqc.collect().ifEmpty([]) +// file ('macs/mergedLibrary/consensus/*') from ch_mlib_macs_consensus_counts_mqc.collect().ifEmpty([]) +// file ('macs/mergedLibrary/consensus/*') from ch_mlib_macs_consensus_deseq_mqc.collect().ifEmpty([]) +// file ('preseq/*') from ch_mlib_preseq_mqc.collect().ifEmpty([]) +// file ('deeptools/*') from ch_mlib_plotprofile_mqc.collect().ifEmpty([]) +// file ('deeptools/*') from ch_mlib_plotfingerprint_mqc.collect().ifEmpty([]) +// +// file ('alignment/mergedReplicate/*') from ch_mrep_bam_flagstat_mqc.collect{it[1]}.ifEmpty([]) +// file ('alignment/mergedReplicate/*') from ch_mrep_bam_stats_mqc.collect().ifEmpty([]) +// file ('alignment/mergedReplicate/*') from ch_mrep_bam_metrics_mqc.collect().ifEmpty([]) +// file ('macs/mergedReplicate/*') from ch_mrep_macs_mqc.collect().ifEmpty([]) +// file ('macs/mergedReplicate/*') from ch_mrep_peak_qc_mqc.collect().ifEmpty([]) +// file ('macs/mergedReplicate/consensus/*') from ch_mrep_macs_consensus_counts_mqc.collect().ifEmpty([]) +// file ('macs/mergedReplicate/consensus/*') from ch_mrep_macs_consensus_deseq_mqc.collect().ifEmpty([]) +// +// output: +// file "*multiqc_report.html" into ch_multiqc_report +// file "*_data" +// file "multiqc_plots" +// +// script: +// rtitle = custom_runName ? "--title \"$custom_runName\"" : '' +// rfilename = custom_runName ? "--filename " + custom_runName.replaceAll('\\W','_').replaceAll('_+','_') + "_multiqc_report" : '' +// custom_config_file = params.multiqc_config ? "--config $mqc_custom_config" : '' +// """ +// multiqc . -f $rtitle $rfilename $custom_config_file \\ +// -m custom_content -m fastqc -m cutadapt -m samtools -m picard -m preseq -m featureCounts -m deeptools +// """ +// } +// +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// /* -- -- */ +// /* -- REPORTS/DOCUMENTATION -- */ +// /* -- -- */ +// /////////////////////////////////////////////////////////////////////////////// +// /////////////////////////////////////////////////////////////////////////////// +// +// /* +// * STEP 11 - Output description HTML +// */ +// process output_documentation { +// publishDir "${params.outdir}/Documentation", mode: 'copy' +// +// input: +// file output_docs from ch_output_docs +// +// output: +// file "results_description.html" +// +// script: +// """ +// markdown_to_html.py $output_docs -o results_description.html +// """ +// } +// +// /* +// * Completion e-mail notification +// */ +// workflow.onComplete { +// +// // Set up the e-mail variables +// def subject = "[nf-core/atacseq] Successful: $workflow.runName" +// if (!workflow.success) { +// subject = "[nf-core/atacseq] FAILED: $workflow.runName" +// } +// def email_fields = [:] +// email_fields['version'] = workflow.manifest.version +// email_fields['runName'] = custom_runName ?: workflow.runName +// email_fields['success'] = workflow.success +// email_fields['dateComplete'] = workflow.complete +// email_fields['duration'] = workflow.duration +// email_fields['exitStatus'] = workflow.exitStatus +// email_fields['errorMessage'] = (workflow.errorMessage ?: 'None') +// email_fields['errorReport'] = (workflow.errorReport ?: 'None') +// email_fields['commandLine'] = workflow.commandLine +// email_fields['projectDir'] = workflow.projectDir +// email_fields['summary'] = summary +// email_fields['summary']['Date Started'] = workflow.start +// email_fields['summary']['Date Completed'] = workflow.complete +// email_fields['summary']['Pipeline script file path'] = workflow.scriptFile +// email_fields['summary']['Pipeline script hash ID'] = workflow.scriptId +// if (workflow.repository) email_fields['summary']['Pipeline repository Git URL'] = workflow.repository +// if (workflow.commitId) email_fields['summary']['Pipeline repository Git Commit'] = workflow.commitId +// if (workflow.revision) email_fields['summary']['Pipeline Git branch/tag'] = workflow.revision +// email_fields['summary']['Nextflow Version'] = workflow.nextflow.version +// email_fields['summary']['Nextflow Build'] = workflow.nextflow.build +// email_fields['summary']['Nextflow Compile Timestamp'] = workflow.nextflow.timestamp +// +// // On success try attach the multiqc report +// def mqc_report = null +// try { +// if (workflow.success) { +// mqc_report = ch_multiqc_report.getVal() +// if (mqc_report.getClass() == ArrayList) { +// log.warn "[nf-core/atacseq] Found multiple reports from process 'multiqc', will use only one" +// mqc_report = mqc_report[0] +// } +// } +// } catch (all) { +// log.warn "[nf-core/atacseq] Could not attach MultiQC report to summary email" +// } +// +// // Check if we are only sending emails on failure +// email_address = params.email +// if (!params.email && params.email_on_fail && !workflow.success) { +// email_address = params.email_on_fail +// } +// +// // Render the TXT template +// def engine = new groovy.text.GStringTemplateEngine() +// def tf = new File("$baseDir/assets/email_template.txt") +// def txt_template = engine.createTemplate(tf).make(email_fields) +// def email_txt = txt_template.toString() +// +// // Render the HTML template +// def hf = new File("$baseDir/assets/email_template.html") +// def html_template = engine.createTemplate(hf).make(email_fields) +// def email_html = html_template.toString() +// +// // Render the sendmail template +// def smail_fields = [ email: email_address, subject: subject, email_txt: email_txt, email_html: email_html, baseDir: "$baseDir", mqcFile: mqc_report, mqcMaxSize: params.max_multiqc_email_size.toBytes() ] +// def sf = new File("$baseDir/assets/sendmail_template.txt") +// def sendmail_template = engine.createTemplate(sf).make(smail_fields) +// def sendmail_html = sendmail_template.toString() +// +// // Send the HTML e-mail +// if (email_address) { +// try { +// if (params.plaintext_email) { throw GroovyException('Send plaintext e-mail, not HTML') } +// // Try to send HTML e-mail using sendmail +// [ 'sendmail', '-t' ].execute() << sendmail_html +// log.info "[nf-core/atacseq] Sent summary e-mail to $email_address (sendmail)" +// } catch (all) { +// // Catch failures and try with plaintext +// [ 'mail', '-s', subject, email_address ].execute() << email_txt +// log.info "[nf-core/atacseq] Sent summary e-mail to $email_address (mail)" +// } +// } +// +// // Write summary e-mail HTML to a file +// def output_d = new File("${params.outdir}/pipeline_info/") +// if (!output_d.exists()) { +// output_d.mkdirs() +// } +// def output_hf = new File(output_d, "pipeline_report.html") +// output_hf.withWriter { w -> w << email_html } +// def output_tf = new File(output_d, "pipeline_report.txt") +// output_tf.withWriter { w -> w << email_txt } +// +// c_green = params.monochrome_logs ? '' : "\033[0;32m"; +// c_purple = params.monochrome_logs ? '' : "\033[0;35m"; +// c_red = params.monochrome_logs ? '' : "\033[0;31m"; +// c_reset = params.monochrome_logs ? '' : "\033[0m"; +// +// if (workflow.stats.ignoredCount > 0 && workflow.success) { +// log.info "-${c_purple}Warning, pipeline completed, but with errored process(es) ${c_reset}-" +// log.info "-${c_red}Number of ignored errored process(es) : ${workflow.stats.ignoredCount} ${c_reset}-" +// log.info "-${c_green}Number of successfully ran process(es) : ${workflow.stats.succeedCount} ${c_reset}-" +// } +// if (workflow.success) { +// log.info "-${c_purple}[nf-core/atacseq]${c_green} Pipeline completed successfully${c_reset}-" +// } else { +// checkHostname() +// log.info "-${c_purple}[nf-core/atacseq]${c_red} Pipeline completed with errors${c_reset}-" +// } +// +// } /////////////////////////////////////////////////////////////////////////////// /////////////////////////////////////////////////////////////////////////////// From 8a5022bce682a538d85e901512ecb9b50dae9f5b Mon Sep 17 00:00:00 2001 From: drpatelh Date: Mon, 24 Feb 2020 10:21:08 +0000 Subject: [PATCH 107/226] Remove comments --- main.nf | 3262 +++++++++++++++++++++++++++---------------------------- 1 file changed, 1631 insertions(+), 1631 deletions(-) diff --git a/main.nf b/main.nf index 12a67b31..2df57e44 100644 --- a/main.nf +++ b/main.nf @@ -520,1637 +520,1637 @@ process FastQC { } } -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// /* -- -- */ -// /* -- ADAPTER TRIMMING -- */ -// /* -- -- */ -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// -// /* -// * STEP 2 - Trim Galore! -// */ -// if (params.skip_trimming) { -// ch_trimmed_reads = ch_raw_reads_trimgalore -// ch_trimgalore_results_mqc = Channel.empty() -// ch_trimgalore_fastqc_reports_mqc = Channel.empty() -// } else { -// process TrimGalore { -// tag "$name" -// label 'process_high' -// publishDir "${params.outdir}/trim_galore", mode: 'copy', -// saveAs: { filename -> -// if (filename.endsWith(".html")) "fastqc/$filename" -// else if (filename.endsWith(".zip")) "fastqc/zips/$filename" -// else if (filename.endsWith("trimming_report.txt")) "logs/$filename" -// else params.save_trimmed ? filename : null -// } -// -// input: -// set val(name), file(reads) from ch_raw_reads_trimgalore -// -// output: -// set val(name), file("*.fq.gz") into ch_trimmed_reads -// file "*.txt" into ch_trimgalore_results_mqc -// file "*.{zip,html}" into ch_trimgalore_fastqc_reports_mqc -// -// script: -// // Calculate number of --cores for TrimGalore based on value of task.cpus -// // See: https://github.com/FelixKrueger/TrimGalore/blob/master/Changelog.md#version-060-release-on-1-mar-2019 -// // See: https://github.com/nf-core/atacseq/pull/65 -// def cores = 1 -// if (task.cpus) { -// cores = (task.cpus as int) - 4 -// if (params.single_end) cores = (task.cpus as int) - 3 -// if (cores < 1) cores = 1 -// if (cores > 4) cores = 4 -// } -// -// // Added soft-links to original fastqs for consistent naming in MultiQC -// c_r1 = params.clip_r1 > 0 ? "--clip_r1 ${params.clip_r1}" : '' -// c_r2 = params.clip_r2 > 0 ? "--clip_r2 ${params.clip_r2}" : '' -// tpc_r1 = params.three_prime_clip_r1 > 0 ? "--three_prime_clip_r1 ${params.three_prime_clip_r1}" : '' -// tpc_r2 = params.three_prime_clip_r2 > 0 ? "--three_prime_clip_r2 ${params.three_prime_clip_r2}" : '' -// nextseq = params.trim_nextseq > 0 ? "--nextseq ${params.trim_nextseq}" : '' -// -// // Added soft-links to original fastqs for consistent naming in MultiQC -// if (params.single_end) { -// """ -// [ ! -f ${name}.fastq.gz ] && ln -s $reads ${name}.fastq.gz -// trim_galore --cores $cores --fastqc --gzip $c_r1 $tpc_r1 $nextseq ${name}.fastq.gz -// """ -// } else { -// """ -// [ ! -f ${name}_1.fastq.gz ] && ln -s ${reads[0]} ${name}_1.fastq.gz -// [ ! -f ${name}_2.fastq.gz ] && ln -s ${reads[1]} ${name}_2.fastq.gz -// trim_galore --cores $cores --paired --fastqc --gzip $c_r1 $c_r2 $tpc_r1 $tpc_r2 $nextseq ${name}_1.fastq.gz ${name}_2.fastq.gz -// """ -// } -// } -// } -// -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// /* -- -- */ -// /* -- ALIGN -- */ -// /* -- -- */ -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// -// /* -// * STEP 3.1 - Align read 1 with bwa -// */ -// process BWAMem { -// tag "$name" -// label 'process_high' -// -// input: -// set val(name), file(reads) from ch_trimmed_reads -// file index from ch_bwa_index.collect() -// -// output: -// set val(name), file("*.bam") into ch_bwa_bam -// -// script: -// prefix = "${name}.Lb" -// rg = "\'@RG\\tID:${name}\\tSM:${name.split('_')[0..-2].join('_')}\\tPL:ILLUMINA\\tLB:${name}\\tPU:1\'" -// if (params.seq_center) { -// rg = "\'@RG\\tID:${name}\\tSM:${name.split('_')[0..-2].join('_')}\\tPL:ILLUMINA\\tLB:${name}\\tPU:1\\tCN:${params.seq_center}\'" -// } -// """ -// bwa mem \\ -// -t $task.cpus \\ -// -M \\ -// -R $rg \\ -// ${index}/${bwa_base} \\ -// $reads \\ -// | samtools view -@ $task.cpus -b -h -F 0x0100 -O BAM -o ${prefix}.bam - -// """ -// } -// -// /* -// * STEP 3.2 - Convert .bam to coordinate sorted .bam -// */ -// process SortBAM { -// tag "$name" -// label 'process_medium' -// if (params.save_align_intermeds) { -// publishDir path: "${params.outdir}/bwa/library", mode: 'copy', -// saveAs: { filename -> -// if (filename.endsWith(".flagstat")) "samtools_stats/$filename" -// else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" -// else if (filename.endsWith(".stats")) "samtools_stats/$filename" -// else filename -// } -// } -// -// input: -// set val(name), file(bam) from ch_bwa_bam -// -// output: -// set val(name), file("*.sorted.{bam,bam.bai}") into ch_sort_bam_merge -// file "*.{flagstat,idxstats,stats}" into ch_sort_bam_flagstat_mqc -// -// script: -// prefix = "${name}.Lb" -// """ -// samtools sort -@ $task.cpus -o ${prefix}.sorted.bam -T $name $bam -// samtools index ${prefix}.sorted.bam -// samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat -// samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats -// samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats -// """ -// } -// -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// /* -- -- */ -// /* -- MERGE LIBRARY BAM -- */ -// /* -- -- */ -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// -// /* -// * STEP 4.1 Merge BAM files for all libraries from same replicate -// */ -// ch_sort_bam_merge -// .map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] } -// .groupTuple(by: [0]) -// .map { it -> [ it[0], it[1].flatten() ] } -// .set { ch_sort_bam_merge } -// -// process MergedLibBAM { -// tag "$name" -// label 'process_medium' -// publishDir "${params.outdir}/bwa/mergedLibrary", mode: 'copy', -// saveAs: { filename -> -// if (filename.endsWith(".flagstat")) "samtools_stats/$filename" -// else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" -// else if (filename.endsWith(".stats")) "samtools_stats/$filename" -// else if (filename.endsWith(".metrics.txt")) "picard_metrics/$filename" -// else params.save_align_intermeds ? filename : null -// } -// -// input: -// set val(name), file(bams) from ch_sort_bam_merge -// -// output: -// set val(name), file("*${prefix}.sorted.{bam,bam.bai}") into ch_mlib_bam_filter, -// ch_mlib_bam_preseq, -// ch_mlib_bam_ataqv -// file "*.{flagstat,idxstats,stats}" into ch_mlib_bam_stats_mqc -// file "*.txt" into ch_mlib_bam_metrics_mqc -// -// script: -// prefix = "${name}.mLb.mkD" -// bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() -// def avail_mem = 3 -// if (!task.memory) { -// log.info "[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." -// } else { -// avail_mem = task.memory.toGiga() -// } -// if (bam_files.size() > 1) { -// """ -// picard -Xmx${avail_mem}g MergeSamFiles \\ -// ${'INPUT='+bam_files.join(' INPUT=')} \\ -// OUTPUT=${name}.sorted.bam \\ -// SORT_ORDER=coordinate \\ -// VALIDATION_STRINGENCY=LENIENT \\ -// TMP_DIR=tmp -// samtools index ${name}.sorted.bam -// -// picard -Xmx${avail_mem}g MarkDuplicates \\ -// INPUT=${name}.sorted.bam \\ -// OUTPUT=${prefix}.sorted.bam \\ -// ASSUME_SORTED=true \\ -// REMOVE_DUPLICATES=false \\ -// METRICS_FILE=${prefix}.MarkDuplicates.metrics.txt \\ -// VALIDATION_STRINGENCY=LENIENT \\ -// TMP_DIR=tmp -// -// samtools index ${prefix}.sorted.bam -// samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats -// samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat -// samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats -// """ -// } else { -// """ -// picard -Xmx${avail_mem}g MarkDuplicates \\ -// INPUT=${bam_files[0]} \\ -// OUTPUT=${prefix}.sorted.bam \\ -// ASSUME_SORTED=true \\ -// REMOVE_DUPLICATES=false \\ -// METRICS_FILE=${prefix}.MarkDuplicates.metrics.txt \\ -// VALIDATION_STRINGENCY=LENIENT \\ -// TMP_DIR=tmp -// -// samtools index ${prefix}.sorted.bam -// samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats -// samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat -// samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats -// """ -// } -// } -// -// /* -// * STEP 4.2 Filter BAM file at merged library-level -// */ -// process MergedLibBAMFilter { -// tag "$name" -// label 'process_medium' -// publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', -// saveAs: { filename -> -// if (params.single_end || params.save_align_intermeds) { -// if (filename.endsWith(".flagstat")) "samtools_stats/$filename" -// else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" -// else if (filename.endsWith(".stats")) "samtools_stats/$filename" -// else if (filename.endsWith(".sorted.bam")) filename -// else if (filename.endsWith(".sorted.bam.bai")) filename -// else null -// } -// } -// -// input: -// set val(name), file(bam) from ch_mlib_bam_filter -// file bed from ch_genome_filter_regions.collect() -// file bamtools_filter_config from ch_bamtools_filter_config -// -// output: -// set val(name), file("*.{bam,bam.bai}") into ch_mlib_filter_bam -// set val(name), file("*.flagstat") into ch_mlib_filter_bam_flagstat -// file "*.{idxstats,stats}" into ch_mlib_filter_bam_stats_mqc -// -// script: -// prefix = params.single_end ? "${name}.mLb.clN" : "${name}.mLb.flT" -// filter_params = params.single_end ? "-F 0x004" : "-F 0x004 -F 0x0008 -f 0x001" -// dup_params = params.keep_dups ? "" : "-F 0x0400" -// multimap_params = params.keep_multi_map ? "" : "-q 1" -// blacklist_params = params.blacklist ? "-L $bed" : "" -// name_sort_bam = params.single_end ? "" : "samtools sort -n -@ $task.cpus -o ${prefix}.bam -T $prefix ${prefix}.sorted.bam" -// """ -// samtools view \\ -// $filter_params \\ -// $dup_params \\ -// $multimap_params \\ -// $blacklist_params \\ -// -b ${bam[0]} \\ -// | bamtools filter \\ -// -out ${prefix}.sorted.bam \\ -// -script $bamtools_filter_config -// -// samtools index ${prefix}.sorted.bam -// samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat -// samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats -// samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats -// -// $name_sort_bam -// """ -// } -// -// /* -// * STEP 4.3 Remove orphan reads from paired-end BAM file -// */ -// if (params.single_end) { -// ch_mlib_filter_bam -// .into { ch_mlib_rm_orphan_bam_metrics; -// ch_mlib_rm_orphan_bam_bigwig; -// ch_mlib_rm_orphan_bam_macs; -// ch_mlib_rm_orphan_bam_plotfingerprint; -// ch_mlib_rm_orphan_bam_mrep; -// ch_mlib_name_bam_mlib_counts; -// ch_mlib_name_bam_mrep_counts } -// -// ch_mlib_filter_bam_flagstat -// .into { ch_mlib_rm_orphan_flagstat_bigwig; -// ch_mlib_rm_orphan_flagstat_macs; -// ch_mlib_rm_orphan_flagstat_mqc } -// -// ch_mlib_filter_bam_stats_mqc -// .set { ch_mlib_rm_orphan_stats_mqc } -// } else { -// process MergedLibBAMRemoveOrphan { -// tag "$name" -// label 'process_medium' -// publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', -// saveAs: { filename -> -// if (filename.endsWith(".flagstat")) "samtools_stats/$filename" -// else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" -// else if (filename.endsWith(".stats")) "samtools_stats/$filename" -// else if (filename.endsWith(".sorted.bam")) filename -// else if (filename.endsWith(".sorted.bam.bai")) filename -// else null -// } -// -// input: -// set val(name), file(bam) from ch_mlib_filter_bam -// -// output: -// set val(name), file("*.sorted.{bam,bam.bai}") into ch_mlib_rm_orphan_bam_metrics, -// ch_mlib_rm_orphan_bam_bigwig, -// ch_mlib_rm_orphan_bam_macs, -// ch_mlib_rm_orphan_bam_plotfingerprint, -// ch_mlib_rm_orphan_bam_mrep -// set val(name), file("${prefix}.bam") into ch_mlib_name_bam_mlib_counts, -// ch_mlib_name_bam_mrep_counts -// set val(name), file("*.flagstat") into ch_mlib_rm_orphan_flagstat_bigwig, -// ch_mlib_rm_orphan_flagstat_macs, -// ch_mlib_rm_orphan_flagstat_mqc -// file "*.{idxstats,stats}" into ch_mlib_rm_orphan_stats_mqc -// -// script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ -// prefix = "${name}.mLb.clN" -// """ -// bampe_rm_orphan.py ${bam[0]} ${prefix}.bam --only_fr_pairs -// -// samtools sort -@ $task.cpus -o ${prefix}.sorted.bam -T $prefix ${prefix}.bam -// samtools index ${prefix}.sorted.bam -// samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat -// samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats -// samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats -// """ -// } -// } -// -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// /* -- -- */ -// /* -- MERGE LIBRARY BAM POST-ANALYSIS -- */ -// /* -- -- */ -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// -// /* -// * STEP 5.1 preseq analysis after merging libraries and before filtering -// */ -// process MergedLibPreseq { -// tag "$name" -// label 'process_low' -// publishDir "${params.outdir}/bwa/mergedLibrary/preseq", mode: 'copy' -// -// when: -// !params.skip_preseq -// -// input: -// set val(name), file(bam) from ch_mlib_bam_preseq -// -// output: -// file "*.ccurve.txt" into ch_mlib_preseq_mqc -// -// script: -// prefix = "${name}.mLb.mkD" -// """ -// preseq lc_extrap -v -output ${prefix}.ccurve.txt -bam ${bam[0]} -// """ -// } -// -// /* -// * STEP 5.2 Picard CollectMultipleMetrics after merging libraries and filtering -// */ -// process MergedLibMetrics { -// tag "$name" -// label 'process_medium' -// publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', -// saveAs: { filename -> -// if (filename.endsWith("_metrics")) "picard_metrics/$filename" -// else if (filename.endsWith(".pdf")) "picard_metrics/pdf/$filename" -// else null -// } -// -// when: -// !params.skip_picard_metrics -// -// input: -// set val(name), file(bam) from ch_mlib_rm_orphan_bam_metrics -// file fasta from ch_fasta -// -// output: -// file "*_metrics" into ch_mlib_collectmetrics_mqc -// file "*.pdf" into ch_mlib_collectmetrics_pdf -// -// script: -// prefix = "${name}.mLb.clN" -// def avail_mem = 3 -// if (!task.memory) { -// log.info "[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." -// } else { -// avail_mem = task.memory.toGiga() -// } -// """ -// picard -Xmx${avail_mem}g CollectMultipleMetrics \\ -// INPUT=${bam[0]} \\ -// OUTPUT=${prefix}.CollectMultipleMetrics \\ -// REFERENCE_SEQUENCE=$fasta \\ -// VALIDATION_STRINGENCY=LENIENT \\ -// TMP_DIR=tmp -// """ -// } -// -// /* -// * STEP 5.3 Read depth normalised bigWig -// */ -// process MergedLibBigWig { -// tag "$name" -// label 'process_medium' -// publishDir "${params.outdir}/bwa/mergedLibrary/bigwig", mode: 'copy', -// saveAs: { filename -> -// if (filename.endsWith("scale_factor.txt")) "scale/$filename" -// else if (filename.endsWith(".bigWig")) "$filename" -// else null -// } -// -// input: -// set val(name), file(bam), file(flagstat) from ch_mlib_rm_orphan_bam_bigwig.join(ch_mlib_rm_orphan_flagstat_bigwig, by: [0]) -// file sizes from ch_genome_sizes_mlib_bigwig.collect() -// -// output: -// set val(name), file("*.bigWig") into ch_mlib_bigwig_plotprofile -// file "*scale_factor.txt" into ch_mlib_bigwig_scale -// file "*igv.txt" into ch_mlib_bigwig_igv -// -// script: -// prefix = "${name}.mLb.clN" -// pe_fragment = params.single_end ? "" : "-pc" -// extend = (params.single_end && params.fragment_size > 0) ? "-fs ${params.fragment_size}" : '' -// """ -// SCALE_FACTOR=\$(grep 'mapped (' $flagstat | awk '{print 1000000/\$1}') -// echo \$SCALE_FACTOR > ${prefix}.scale_factor.txt -// genomeCoverageBed -ibam ${bam[0]} -bg -scale \$SCALE_FACTOR $pe_fragment $extend | sort -T '.' -k1,1 -k2,2n > ${prefix}.bedGraph -// -// bedGraphToBigWig ${prefix}.bedGraph $sizes ${prefix}.bigWig -// -// find * -type f -name "*.bigWig" -exec echo -e "bwa/mergedLibrary/bigwig/"{}"\\t0,0,178" \\; > ${prefix}.bigWig.igv.txt -// """ -// } -// -// /* -// * STEP 5.4 generate gene body coverage plot with deepTools -// */ -// process MergedLibPlotProfile { -// tag "$name" -// label 'process_high' -// publishDir "${params.outdir}/bwa/mergedLibrary/deepTools/plotProfile", mode: 'copy' -// -// when: -// !params.skip_plot_profile -// -// input: -// set val(name), file(bigwig) from ch_mlib_bigwig_plotprofile -// file bed from ch_gene_bed -// -// output: -// file '*.{gz,pdf}' into ch_mlib_plotprofile_results -// file '*.plotProfile.tab' into ch_mlib_plotprofile_mqc -// -// script: -// prefix = "${name}.mLb.clN" -// """ -// computeMatrix scale-regions \\ -// --regionsFileName $bed \\ -// --scoreFileName $bigwig \\ -// --outFileName ${prefix}.computeMatrix.mat.gz \\ -// --outFileNameMatrix ${prefix}.computeMatrix.vals.mat.gz \\ -// --regionBodyLength 1000 \\ -// --beforeRegionStartLength 3000 \\ -// --afterRegionStartLength 3000 \\ -// --skipZeros \\ -// --smartLabels \\ -// --numberOfProcessors $task.cpus -// -// plotProfile --matrixFile ${prefix}.computeMatrix.mat.gz \\ -// --outFileName ${prefix}.plotProfile.pdf \\ -// --outFileNameData ${prefix}.plotProfile.tab -// """ -// } -// -// /* -// * STEP 5.5 deepTools plotFingerprint -// */ -// process MergedLibPlotFingerprint { -// tag "$name" -// label 'process_high' -// publishDir "${params.outdir}/bwa/mergedLibrary/deepTools/plotFingerprint", mode: 'copy' -// -// when: -// !params.skip_plot_fingerprint -// -// input: -// set val(name), file(bam) from ch_mlib_rm_orphan_bam_plotfingerprint -// -// output: -// file '*.{txt,pdf}' into ch_mlib_plotfingerprint_results -// file '*.raw.txt' into ch_mlib_plotfingerprint_mqc -// -// script: -// prefix = "${name}.mLb.clN" -// extend = (params.single_end && params.fragment_size > 0) ? "--extendReads ${params.fragment_size}" : '' -// """ -// plotFingerprint \\ -// --bamfiles ${bam[0]} \\ -// --plotFile ${prefix}.plotFingerprint.pdf \\ -// $extend \\ -// --labels $prefix \\ -// --outRawCounts ${prefix}.plotFingerprint.raw.txt \\ -// --outQualityMetrics ${prefix}.plotFingerprint.qcmetrics.txt \\ -// --skipZeros \\ -// --numberOfProcessors $task.cpus \\ -// --numberOfSamples $params.fingerprint_bins -// """ -// } -// -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// /* -- -- */ -// /* -- MERGE LIBRARY PEAK ANALYSIS -- */ -// /* -- -- */ -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// -// /* -// * STEP 6.1 Call peaks with MACS2 and calculate FRiP score -// */ -// process MergedLibMACSCallPeak { -// tag "$name" -// label 'process_medium' -// publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}", mode: 'copy', -// saveAs: { filename -> -// if (filename.endsWith(".tsv")) "qc/$filename" -// else if (filename.endsWith(".igv.txt")) null -// else filename -// } -// -// when: -// params.macs_gsize -// -// input: -// set val(name), file(bam), file(flagstat) from ch_mlib_rm_orphan_bam_macs.join(ch_mlib_rm_orphan_flagstat_macs, by: [0]) -// file mlib_peak_count_header from ch_mlib_peak_count_header -// file mlib_frip_score_header from ch_mlib_frip_score_header -// -// output: -// set val(name), file("*.{bed,xls,gappedPeak,bdg}") into ch_mlib_macs_output -// set val(name), file("*$PEAK_TYPE") into ch_mlib_macs_homer, -// ch_mlib_macs_qc, -// ch_mlib_macs_consensus, -// ch_mlib_macs_ataqv -// file "*igv.txt" into ch_mlib_macs_igv -// file "*_mqc.tsv" into ch_mlib_macs_mqc -// -// script: -// prefix = "${name}.mLb.clN" -// broad = params.narrow_peak ? '' : "--broad --broad-cutoff ${params.broad_cutoff}" -// format = params.single_end ? "BAM" : "BAMPE" -// pileup = params.save_macs_pileup ? "-B --SPMR" : "" -// """ -// macs2 callpeak \\ -// -t ${bam[0]} \\ -// $broad \\ -// -f $format \\ -// -g $params.macs_gsize \\ -// -n $prefix \\ -// $pileup \\ -// --keep-dup all \\ -// --nomodel -// -// cat ${prefix}_peaks.${PEAK_TYPE} | wc -l | awk -v OFS='\t' '{ print "${name}", \$1 }' | cat $mlib_peak_count_header - > ${prefix}_peaks.count_mqc.tsv -// -// READS_IN_PEAKS=\$(intersectBed -a ${bam[0]} -b ${prefix}_peaks.${PEAK_TYPE} -bed -c -f 0.20 | awk -F '\t' '{sum += \$NF} END {print sum}') -// grep 'mapped (' $flagstat | awk -v a="\$READS_IN_PEAKS" -v OFS='\t' '{print "${name}", a/\$1}' | cat $mlib_frip_score_header - > ${prefix}_peaks.FRiP_mqc.tsv -// -// find * -type f -name "*.${PEAK_TYPE}" -exec echo -e "bwa/mergedLibrary/macs/${PEAK_TYPE}/"{}"\\t0,0,178" \\; > ${prefix}_peaks.igv.txt -// """ -// } -// -// /* -// * STEP 6.2 Annotate peaks with HOMER -// */ -// process MergedLibAnnotatePeaks { -// tag "$name" -// label "process_medium" -// publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}", mode: 'copy' -// -// when: -// params.macs_gsize -// -// input: -// set val(name), file(peak) from ch_mlib_macs_homer -// file fasta from ch_fasta -// file gtf from ch_gtf -// -// output: -// file "*.txt" into ch_mlib_macs_annotate -// -// script: -// prefix = "${name}.mLb.clN" -// """ -// annotatePeaks.pl \\ -// $peak \\ -// $fasta \\ -// -gid \\ -// -gtf $gtf \\ -// -cpu $task.cpus \\ -// > ${prefix}_peaks.annotatePeaks.txt -// """ -// } -// -// /* -// * STEP 6.3 Aggregated QC plots for peaks, FRiP and peak-to-gene annotation -// */ -// process MergedLibPeakQC { -// label "process_medium" -// publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/qc", mode: 'copy' -// -// when: -// params.macs_gsize -// -// input: -// file peaks from ch_mlib_macs_qc.collect{ it[1] } -// file annos from ch_mlib_macs_annotate.collect() -// file mlib_peak_annotation_header from ch_mlib_peak_annotation_header -// -// output: -// file "*.{txt,pdf}" into ch_mlib_peak_qc -// file "*.tsv" into ch_mlib_peak_qc_mqc -// -// script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ -// suffix = 'mLb.clN' -// """ -// plot_macs_qc.r \\ -// -i ${peaks.join(',')} \\ -// -s ${peaks.join(',').replaceAll(".${suffix}_peaks.${PEAK_TYPE}","")} \\ -// -o ./ \\ -// -p macs_peak.${suffix} -// -// plot_homer_annotatepeaks.r \\ -// -i ${annos.join(',')} \\ -// -s ${annos.join(',').replaceAll(".${suffix}_peaks.annotatePeaks.txt","")} \\ -// -o ./ \\ -// -p macs_annotatePeaks.${suffix} -// -// cat $mlib_peak_annotation_header macs_annotatePeaks.${suffix}.summary.txt > macs_annotatePeaks.${suffix}.summary_mqc.tsv -// """ -// } -// -// /* -// * STEP 6.4 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection -// */ -// process MergedLibConsensusPeakSet { -// label 'process_long' -// publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: 'copy', -// saveAs: { filename -> -// if (filename.endsWith(".igv.txt")) null -// else filename -// } -// -// when: -// params.macs_gsize && (replicatesExist || multipleGroups) && !params.skip_consensus_peaks -// -// input: -// file peaks from ch_mlib_macs_consensus.collect{ it[1] } -// -// output: -// file "*.bed" into ch_mlib_macs_consensus_bed -// file "*.saf" into ch_mlib_macs_consensus_saf -// file "*.boolean.txt" into ch_mlib_macs_consensus_bool -// file "*.intersect.{txt,plot.pdf}" into ch_mlib_macs_consensus_intersect -// file "*igv.txt" into ch_mlib_macs_consensus_igv -// -// script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ -// suffix = 'mLb.clN' -// prefix = "consensus_peaks.${suffix}" -// mergecols = params.narrow_peak ? (2..10).join(',') : (2..9).join(',') -// collapsecols = params.narrow_peak ? (["collapse"]*9).join(',') : (["collapse"]*8).join(',') -// expandparam = params.narrow_peak ? "--is_narrow_peak" : "" -// """ -// sort -T '.' -k1,1 -k2,2n ${peaks.collect{it.toString()}.sort().join(' ')} \\ -// | mergeBed -c $mergecols -o $collapsecols > ${prefix}.txt -// -// macs2_merged_expand.py \\ -// ${prefix}.txt \\ -// ${peaks.collect{it.toString()}.sort().join(',').replaceAll("_peaks.${PEAK_TYPE}","")} \\ -// ${prefix}.boolean.txt \\ -// --min_replicates $params.min_reps_consensus \\ -// $expandparam -// -// awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$1, \$2, \$3, \$4, "0", "+" }' ${prefix}.boolean.txt > ${prefix}.bed -// -// echo -e "GeneID\tChr\tStart\tEnd\tStrand" > ${prefix}.saf -// awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$4, \$1, \$2, \$3, "+" }' ${prefix}.boolean.txt >> ${prefix}.saf -// -// sed -i 's/.${suffix}//g' ${prefix}.boolean.intersect.txt -// plot_peak_intersect.r -i ${prefix}.boolean.intersect.txt -o ${prefix}.boolean.intersect.plot.pdf -// -// find * -type f -name "${prefix}.bed" -exec echo -e "bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus/"{}"\\t0,0,0" \\; > ${prefix}.bed.igv.txt -// """ -// } -// -// /* -// * STEP 6.5 Annotate consensus peaks with HOMER, and add annotation to boolean output file -// */ -// process MergedLibConsensusPeakSetAnnotate { -// label "process_medium" -// publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: 'copy' -// -// when: -// params.macs_gsize && (replicatesExist || multipleGroups) && !params.skip_consensus_peaks -// -// input: -// file bed from ch_mlib_macs_consensus_bed -// file bool from ch_mlib_macs_consensus_bool -// file fasta from ch_fasta -// file gtf from ch_gtf -// -// output: -// file "*.annotatePeaks.txt" into ch_mlib_macs_consensus_annotate -// -// script: -// prefix = "consensus_peaks.mLb.clN" -// """ -// annotatePeaks.pl \\ -// $bed \\ -// $fasta \\ -// -gid \\ -// -gtf $gtf \\ -// -cpu $task.cpus \\ -// > ${prefix}.annotatePeaks.txt -// -// cut -f2- ${prefix}.annotatePeaks.txt | awk 'NR==1; NR > 1 {print \$0 | "sort -T '.' -k1,1 -k2,2n"}' | cut -f6- > tmp.txt -// paste $bool tmp.txt > ${prefix}.boolean.annotatePeaks.txt -// """ -// } -// -// /* -// * STEP 6.6 Count reads in consensus peaks with featureCounts -// */ -// process MergedLibConsensusPeakSetCounts { -// label 'process_medium' -// publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: 'copy' -// -// when: -// params.macs_gsize && (replicatesExist || multipleGroups) && !params.skip_consensus_peaks -// -// input: -// file bams from ch_mlib_name_bam_mlib_counts.collect{ it[1] } -// file saf from ch_mlib_macs_consensus_saf.collect() -// -// output: -// file "*featureCounts.txt" into ch_mlib_macs_consensus_counts -// file "*featureCounts.txt.summary" into ch_mlib_macs_consensus_counts_mqc -// -// script: -// prefix = "consensus_peaks.mLb.clN" -// bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() -// pe_params = params.single_end ? '' : "-p --donotsort" -// """ -// featureCounts \\ -// -F SAF \\ -// -O \\ -// --fracOverlap 0.2 \\ -// -T $task.cpus \\ -// $pe_params \\ -// -a $saf \\ -// -o ${prefix}.featureCounts.txt \\ -// ${bam_files.join(' ')} -// """ -// } -// -// /* -// * STEP 6.7 Perform differential analysis with DESeq2 -// */ -// process MergedLibConsensusPeakSetDESeq { -// label 'process_medium' -// publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus/deseq2", mode: 'copy', -// saveAs: { filename -> -// if (filename.endsWith(".igv.txt")) null -// else filename -// } -// -// when: -// params.macs_gsize && replicatesExist && multipleGroups && !params.skip_consensus_peaks && !params.skip_diff_analysis -// -// input: -// file counts from ch_mlib_macs_consensus_counts -// file mlib_deseq2_pca_header from ch_mlib_deseq2_pca_header -// file mlib_deseq2_clustering_header from ch_mlib_deseq2_clustering_header -// -// output: -// file "*.{RData,results.txt,pdf,log}" into ch_mlib_macs_consensus_deseq_results -// file "sizeFactors" into ch_mlib_macs_consensus_deseq_factors -// file "*vs*/*.{pdf,txt}" into ch_mlib_macs_consensus_deseq_comp_results -// file "*vs*/*.bed" into ch_mlib_macs_consensus_deseq_comp_bed -// file "*igv.txt" into ch_mlib_macs_consensus_deseq_comp_igv -// file "*.tsv" into ch_mlib_macs_consensus_deseq_mqc -// -// script: -// prefix = "consensus_peaks.mLb.clN" -// bam_ext = params.single_end ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" -// """ -// featurecounts_deseq2.r \\ -// --featurecount_file $counts \\ -// --bam_suffix '$bam_ext' \\ -// --outdir ./ \\ -// --outprefix $prefix \\ -// --outsuffix .mLb \\ -// --cores $task.cpus -// -// cat $mlib_deseq2_pca_header ${prefix}.pca.vals.txt > ${prefix}.pca.vals_mqc.tsv -// cat $mlib_deseq2_clustering_header ${prefix}.sample.dists.txt > ${prefix}.sample.dists_mqc.tsv -// -// find * -type f -name "*.FDR0.05.results.bed" -exec echo -e "bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus/deseq2/"{}"\\t255,0,0" \\; > ${prefix}.igv.txt -// """ -// } -// -// /* -// * STEP 6.8 Run ataqv on BAM file and corresponding peaks -// */ -// process MergedLibAtaqv { -// tag "$name" -// label 'process_medium' -// publishDir "${params.outdir}/bwa/mergedLibrary/ataqv/${PEAK_TYPE}", mode: 'copy' -// -// when: -// !params.skip_ataqv -// -// input: -// set val(name), file(bam), file(peak) from ch_mlib_bam_ataqv.join(ch_mlib_macs_ataqv, by: [0]) -// file autosomes from ch_genome_autosomes.collect() -// file tss_bed from ch_tss_bed -// -// output: -// file "*.json" into ch_mlib_ataqv -// -// script: -// peak_param = params.macs_gsize ? "--peak-file ${peak}" : "" -// mito_param = params.mito_name ? "--mitochondrial-reference-name ${params.mito_name}" : "" -// """ -// ataqv \\ -// --threads $task.cpus \\ -// $peak_param \\ -// --tss-file $tss_bed \\ -// --metrics-file ${name}.ataqv.json \\ -// --name $name \\ -// --ignore-read-groups \\ -// --autosomal-reference-file $autosomes \\ -// $mito_param \\ -// NA \\ -// ${bam[0]} -// """ -// } -// -// /* -// * STEP 6.9 run ataqv mkarv on all JSON files to render web app -// */ -// process MergedLibAtaqvMkarv { -// label 'process_medium' -// publishDir "${params.outdir}/bwa/mergedLibrary/ataqv/${PEAK_TYPE}", mode: 'copy' -// -// when: -// !params.skip_ataqv -// -// input: -// file json from ch_mlib_ataqv.collect() -// -// output: -// file "html" into ch_mlib_ataqv_mkarv -// -// script: -// """ -// mkarv \\ -// --concurrency $task.cpus \\ -// --force \\ -// ./html/ \\ -// ${json.join(' ')} -// """ -// } -// -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// /* -- -- */ -// /* -- MERGE REPLICATE BAM -- */ -// /* -- -- */ -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// -// /* -// * STEP 7 Merge library BAM files across all replicates -// */ -// ch_mlib_rm_orphan_bam_mrep -// .map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] } -// .groupTuple(by: [0]) -// .map { it -> [ it[0], it[1].flatten() ] } -// .set { ch_mlib_rm_orphan_bam_mrep } -// -// process MergedRepBAM { -// tag "$name" -// label 'process_medium' -// publishDir "${params.outdir}/bwa/mergedReplicate", mode: 'copy', -// saveAs: { filename -> -// if (filename.endsWith(".flagstat")) "samtools_stats/$filename" -// else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" -// else if (filename.endsWith(".stats")) "samtools_stats/$filename" -// else if (filename.endsWith(".metrics.txt")) "picard_metrics/$filename" -// else filename -// } -// -// input: -// set val(name), file(bams) from ch_mlib_rm_orphan_bam_mrep -// -// output: -// set val(name), file("*${prefix}.sorted.{bam,bam.bai}") into ch_mrep_bam_bigwig, -// ch_mrep_bam_macs -// set val(name), file("*.flagstat") into ch_mrep_bam_flagstat_bigwig, -// ch_mrep_bam_flagstat_macs, -// ch_mrep_bam_flagstat_mqc -// file "*.{idxstats,stats}" into ch_mrep_bam_stats_mqc -// file "*.txt" into ch_mrep_bam_metrics_mqc -// -// when: -// !params.skip_merge_replicates && replicatesExist -// -// script: -// prefix = "${name}.mRp.clN" -// bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() -// def avail_mem = 3 -// if (!task.memory) { -// log.info "[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." -// } else { -// avail_mem = task.memory.toGiga() -// } -// if (bam_files.size() > 1) { -// """ -// picard -Xmx${avail_mem}g MergeSamFiles \\ -// ${'INPUT='+bam_files.join(' INPUT=')} \\ -// OUTPUT=${name}.sorted.bam \\ -// SORT_ORDER=coordinate \\ -// VALIDATION_STRINGENCY=LENIENT \\ -// TMP_DIR=tmp -// samtools index ${name}.sorted.bam -// -// picard -Xmx${avail_mem}g MarkDuplicates \\ -// INPUT=${name}.sorted.bam \\ -// OUTPUT=${prefix}.sorted.bam \\ -// ASSUME_SORTED=true \\ -// REMOVE_DUPLICATES=true \\ -// METRICS_FILE=${prefix}.MarkDuplicates.metrics.txt \\ -// VALIDATION_STRINGENCY=LENIENT \\ -// TMP_DIR=tmp -// -// samtools index ${prefix}.sorted.bam -// samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat -// samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats -// samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats -// """ -// } else { -// """ -// ln -s ${bams[0]} ${prefix}.sorted.bam -// ln -s ${bams[1]} ${prefix}.sorted.bam.bai -// touch ${prefix}.MarkDuplicates.metrics.txt -// samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat -// samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats -// samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats -// """ -// } -// } -// -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// /* -- -- */ -// /* -- MERGE REPLICATE BAM POST-ANALYSIS -- */ -// /* -- -- */ -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// -// /* -// * STEP 8.1 Read depth normalised bigWig -// */ -// process MergedRepBigWig { -// tag "$name" -// label 'process_medium' -// publishDir "${params.outdir}/bwa/mergedReplicate/bigwig", mode: 'copy', -// saveAs: { filename -> -// if (filename.endsWith("scale_factor.txt")) "scale/$filename" -// else if (filename.endsWith(".bigWig")) "$filename" -// else null -// } -// -// when: -// !params.skip_merge_replicates && replicatesExist -// -// input: -// set val(name), file(bam), file(flagstat) from ch_mrep_bam_bigwig.join(ch_mrep_bam_flagstat_bigwig, by: [0]) -// file sizes from ch_genome_sizes_mrep_bigwig.collect() -// -// output: -// set val(name), file("*.bigWig") into ch_mrep_bigwig -// file "*scale_factor.txt" into ch_mrep_bigwig_scale -// file "*igv.txt" into ch_mrep_bigwig_igv -// -// script: -// prefix = "${name}.mRp.clN" -// pe_fragment = params.single_end ? "" : "-pc" -// extend = (params.single_end && params.fragment_size > 0) ? "-fs ${params.fragment_size}" : '' -// """ -// SCALE_FACTOR=\$(grep 'mapped (' $flagstat | awk '{print 1000000/\$1}') -// echo \$SCALE_FACTOR > ${prefix}.scale_factor.txt -// genomeCoverageBed -ibam ${bam[0]} -bg -scale \$SCALE_FACTOR $pe_fragment $extend | sort -T '.' -k1,1 -k2,2n > ${prefix}.bedGraph -// -// bedGraphToBigWig ${prefix}.bedGraph $sizes ${prefix}.bigWig -// -// find * -type f -name "*.bigWig" -exec echo -e "bwa/mergedReplicate/bigwig/"{}"\\t0,0,178" \\; > ${prefix}.bigWig.igv.txt -// """ -// } -// -// /* -// * STEP 8.2 Call peaks with MACS2 and calculate FRiP score -// */ -// process MergedRepMACSCallPeak { -// tag "$name" -// label 'process_medium' -// publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}", mode: 'copy', -// saveAs: { filename -> -// if (filename.endsWith(".tsv")) "qc/$filename" -// else if (filename.endsWith(".igv.txt")) null -// else filename -// } -// -// when: -// !params.skip_merge_replicates && replicatesExist && params.macs_gsize -// -// input: -// set val(name), file(bam), file(flagstat) from ch_mrep_bam_macs.join(ch_mrep_bam_flagstat_macs, by: [0]) -// file mrep_peak_count_header from ch_mrep_peak_count_header -// file mrep_frip_score_header from ch_mrep_frip_score_header -// -// output: -// set val(name), file("*.{bed,xls,gappedPeak,bdg}") into ch_mrep_macs_output -// set val(name), file("*$PEAK_TYPE") into ch_mrep_macs_homer, -// ch_mrep_macs_qc, -// ch_mrep_macs_consensus -// file "*igv.txt" into ch_mrep_macs_igv -// file "*_mqc.tsv" into ch_mrep_macs_mqc -// -// script: -// prefix = "${name}.mRp.clN" -// broad = params.narrow_peak ? '' : "--broad --broad-cutoff ${params.broad_cutoff}" -// format = params.single_end ? "BAM" : "BAMPE" -// pileup = params.save_macs_pileup ? "-B --SPMR" : "" -// """ -// macs2 callpeak \\ -// -t ${bam[0]} \\ -// $broad \\ -// -f $format \\ -// -g $params.macs_gsize \\ -// -n $prefix \\ -// $pileup \\ -// --keep-dup all \\ -// --nomodel -// -// cat ${prefix}_peaks.${PEAK_TYPE} | wc -l | awk -v OFS='\t' '{ print "${name}", \$1 }' | cat $mrep_peak_count_header - > ${prefix}_peaks.count_mqc.tsv -// -// READS_IN_PEAKS=\$(intersectBed -a ${bam[0]} -b ${prefix}_peaks.${PEAK_TYPE} -bed -c -f 0.20 | awk -F '\t' '{sum += \$NF} END {print sum}') -// grep 'mapped (' $flagstat | awk -v a="\$READS_IN_PEAKS" -v OFS='\t' '{print "${name}", a/\$1}' | cat $mrep_frip_score_header - > ${prefix}_peaks.FRiP_mqc.tsv -// -// find * -type f -name "*.${PEAK_TYPE}" -exec echo -e "bwa/mergedReplicate/macs/${PEAK_TYPE}/"{}"\\t0,0,178" \\; > ${prefix}_peaks.igv.txt -// """ -// } -// -// /* -// * STEP 8.3 Annotate peaks with HOMER -// */ -// process MergedRepAnnotatePeaks { -// tag "$name" -// label "process_medium" -// publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}", mode: 'copy' -// -// when: -// !params.skip_merge_replicates && replicatesExist && params.macs_gsize -// -// input: -// set val(name), file(peak) from ch_mrep_macs_homer -// file fasta from ch_fasta -// file gtf from ch_gtf -// -// output: -// file "*.txt" into ch_mrep_macs_annotate -// -// script: -// prefix = "${name}.mRp.clN" -// """ -// annotatePeaks.pl \\ -// $peak \\ -// $fasta \\ -// -gid \\ -// -gtf $gtf \\ -// -cpu $task.cpus \\ -// > ${prefix}_peaks.annotatePeaks.txt -// """ -// } -// -// /* -// * STEP 8.4 Aggregated QC plots for peaks, FRiP and peak-to-gene annotation -// */ -// process MergedRepPeakQC { -// label "process_medium" -// publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/qc", mode: 'copy' -// -// when: -// !params.skip_merge_replicates && replicatesExist && params.macs_gsize -// -// input: -// file peaks from ch_mrep_macs_qc.collect{ it[1] } -// file annos from ch_mrep_macs_annotate.collect() -// file mrep_peak_annotation_header from ch_mrep_peak_annotation_header -// -// output: -// file "*.{txt,pdf}" into ch_mrep_peak_qc -// file "*.tsv" into ch_mrep_peak_qc_mqc -// -// script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ -// suffix = 'mRp.clN' -// """ -// plot_macs_qc.r \\ -// -i ${peaks.join(',')} \\ -// -s ${peaks.join(',').replaceAll(".${suffix}_peaks.${PEAK_TYPE}","")} \\ -// -o ./ \\ -// -p macs_peak.${suffix} -// -// plot_homer_annotatepeaks.r \\ -// -i ${annos.join(',')} \\ -// -s ${annos.join(',').replaceAll(".${suffix}_peaks.annotatePeaks.txt","")} \\ -// -o ./ \\ -// -p macs_annotatePeaks.${suffix} -// -// cat $mrep_peak_annotation_header macs_annotatePeaks.${suffix}.summary.txt > macs_annotatePeaks.${suffix}.summary_mqc.tsv -// """ -// } -// -// /* -// * STEP 8.5 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection -// */ -// process MergedRepConsensusPeakSet { -// label 'process_long' -// publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: 'copy', -// saveAs: { filename -> -// if (filename.endsWith(".igv.txt")) null -// else filename -// } -// -// when: -// !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks -// -// input: -// file peaks from ch_mrep_macs_consensus.collect{ it[1] } -// -// output: -// file "*.bed" into ch_mrep_macs_consensus_bed -// file "*.saf" into ch_mrep_macs_consensus_saf -// file "*.boolean.txt" into ch_mrep_macs_consensus_bool -// file "*.intersect.{txt,plot.pdf}" into ch_mrep_macs_consensus_intersect -// file "*igv.txt" into ch_mrep_macs_consensus_igv -// -// script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ -// suffix = 'mRp.clN' -// prefix = "consensus_peaks.${suffix}" -// mergecols = params.narrow_peak ? (2..10).join(',') : (2..9).join(',') -// collapsecols = params.narrow_peak ? (["collapse"]*9).join(',') : (["collapse"]*8).join(',') -// expandparam = params.narrow_peak ? "--is_narrow_peak" : "" -// """ -// sort -T '.' -k1,1 -k2,2n ${peaks.collect{it.toString()}.sort().join(' ')} \\ -// | mergeBed -c $mergecols -o $collapsecols > ${prefix}.txt -// -// macs2_merged_expand.py \\ -// ${prefix}.txt \\ -// ${peaks.collect{it.toString()}.sort().join(',').replaceAll("_peaks.${PEAK_TYPE}","")} \\ -// ${prefix}.boolean.txt \\ -// $expandparam -// -// awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$1, \$2, \$3, \$4, "0", "+" }' ${prefix}.boolean.txt > ${prefix}.bed -// -// echo -e "GeneID\tChr\tStart\tEnd\tStrand" > ${prefix}.saf -// awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$4, \$1, \$2, \$3, "+" }' ${prefix}.boolean.txt >> ${prefix}.saf -// -// sed -i 's/.${suffix}//g' ${prefix}.boolean.intersect.txt -// plot_peak_intersect.r -i ${prefix}.boolean.intersect.txt -o ${prefix}.boolean.intersect.plot.pdf -// -// find * -type f -name "${prefix}.bed" -exec echo -e "bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus/"{}"\\t0,0,0" \\; > ${prefix}.bed.igv.txt -// """ -// } -// -// /* -// * STEP 8.6 Annotate consensus peaks with HOMER, and add annotation to boolean output file -// */ -// process MergedRepConsensusPeakSetAnnotate { -// label "process_medium" -// publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: 'copy' -// -// when: -// !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks -// -// input: -// file bed from ch_mrep_macs_consensus_bed -// file bool from ch_mrep_macs_consensus_bool -// file fasta from ch_fasta -// file gtf from ch_gtf -// -// output: -// file "*.annotatePeaks.txt" into ch_mrep_macs_consensus_annotate -// -// script: -// prefix = "consensus_peaks.mRp.clN" -// """ -// annotatePeaks.pl \\ -// $bed \\ -// $fasta \\ -// -gid \\ -// -gtf $gtf \\ -// -cpu $task.cpus \\ -// > ${prefix}.annotatePeaks.txt -// -// cut -f2- ${prefix}.annotatePeaks.txt | awk 'NR==1; NR > 1 {print \$0 | "sort -T '.' -k1,1 -k2,2n"}' | cut -f6- > tmp.txt -// paste $bool tmp.txt > ${prefix}.boolean.annotatePeaks.txt -// """ -// } -// -// /* -// * STEP 8.7 Count reads in consensus peaks with featureCounts -// */ -// process MergedRepConsensusPeakSetCounts { -// label 'process_medium' -// publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: 'copy' -// -// when: -// !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks -// -// input: -// file bams from ch_mlib_name_bam_mrep_counts.collect{ it[1] } -// file saf from ch_mrep_macs_consensus_saf.collect() -// -// output: -// file "*featureCounts.txt" into ch_mrep_macs_consensus_counts -// file "*featureCounts.txt.summary" into ch_mrep_macs_consensus_counts_mqc -// -// script: -// prefix = "consensus_peaks.mRp.clN" -// bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() -// pe_params = params.single_end ? '' : "-p --donotsort" -// """ -// featureCounts \\ -// -F SAF \\ -// -O \\ -// --fracOverlap 0.2 \\ -// -T $task.cpus \\ -// $pe_params \\ -// -a $saf \\ -// -o ${prefix}.featureCounts.txt \\ -// ${bam_files.join(' ')} -// """ -// } -// -// /* -// * STEP 8.8 Perform differential analysis with DESeq2 -// */ -// process MergedRepConsensusPeakSetDESeq { -// label 'process_medium' -// publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus/deseq2", mode: 'copy', -// saveAs: { filename -> -// if (filename.endsWith(".igv.txt")) null -// else filename -// } -// -// when: -// !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks && !params.skip_diff_analysis -// -// input: -// file counts from ch_mrep_macs_consensus_counts -// file mrep_deseq2_pca_header from ch_mrep_deseq2_pca_header -// file mrep_deseq2_clustering_header from ch_mrep_deseq2_clustering_header -// -// output: -// file "*.{RData,results.txt,pdf,log}" into ch_mrep_macs_consensus_deseq_results -// file "sizeFactors" into ch_mrep_macs_consensus_deseq_factors -// file "*vs*/*.{pdf,txt}" into ch_mrep_macs_consensus_deseq_comp_results -// file "*vs*/*.bed" into ch_mrep_macs_consensus_deseq_comp_bed -// file "*igv.txt" into ch_mrep_macs_consensus_deseq_comp_igv -// file "*.tsv" into ch_mrep_macs_consensus_deseq_mqc -// -// script: -// prefix = "consensus_peaks.mRp.clN" -// bam_ext = params.single_end ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" -// """ -// featurecounts_deseq2.r \\ -// --featurecount_file $counts \\ -// --bam_suffix '$bam_ext' \\ -// --outdir ./ \\ -// --outprefix $prefix \\ -// --outsuffix .mRp \\ -// --cores $task.cpus -// -// cat $mrep_deseq2_pca_header ${prefix}.pca.vals.txt > ${prefix}.pca.vals_mqc.tsv -// cat $mrep_deseq2_clustering_header ${prefix}.sample.dists.txt > ${prefix}.sample.dists_mqc.tsv -// -// find * -type f -name "*.FDR0.05.results.bed" -exec echo -e "bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus/deseq2/"{}"\\t255,0,0" \\; > ${prefix}.igv.txt -// """ -// } -// -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// /* -- -- */ -// /* -- IGV -- */ -// /* -- -- */ -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// -// /* -// * STEP 9 - Create IGV session file -// */ -// process IGV { -// publishDir "${params.outdir}/igv/${PEAK_TYPE}", mode: 'copy' -// -// when: -// !params.skip_igv -// -// input: -// file fasta from ch_fasta -// -// file bigwigs from ch_mlib_bigwig_igv.collect().ifEmpty([]) -// file peaks from ch_mlib_macs_igv.collect().ifEmpty([]) -// file consensus_peaks from ch_mlib_macs_consensus_igv.collect().ifEmpty([]) -// file differential_peaks from ch_mlib_macs_consensus_deseq_comp_igv.collect().ifEmpty([]) -// -// file rbigwigs from ch_mrep_bigwig_igv.collect().ifEmpty([]) -// file rpeaks from ch_mrep_macs_igv.collect().ifEmpty([]) -// file rconsensus_peaks from ch_mrep_macs_consensus_igv.collect().ifEmpty([]) -// file rdifferential_peaks from ch_mrep_macs_consensus_deseq_comp_igv.collect().ifEmpty([]) -// -// output: -// file "*.{txt,xml}" into ch_igv_session -// -// script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ -// """ -// cat *.txt > igv_files.txt -// igv_files_to_session.py igv_session.xml igv_files.txt ../../genome/${fasta.getName()} --path_prefix '../../' -// """ -// } -// -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// /* -- -- */ -// /* -- MULTIQC -- */ -// /* -- -- */ -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// -// /* -// * Parse software version numbers -// */ -// process get_software_versions { -// publishDir "${params.outdir}/pipeline_info", mode: 'copy', -// saveAs: { filename -> -// if (filename.indexOf(".csv") > 0) filename -// else null -// } -// -// output: -// file 'software_versions_mqc.yaml' into ch_software_versions_mqc -// file "software_versions.csv" -// -// script: -// """ -// echo $workflow.manifest.version > v_pipeline.txt -// echo $workflow.nextflow.version > v_nextflow.txt -// fastqc --version > v_fastqc.txt -// trim_galore --version > v_trim_galore.txt -// echo \$(bwa 2>&1) > v_bwa.txt -// samtools --version > v_samtools.txt -// bedtools --version > v_bedtools.txt -// echo \$(bamtools --version 2>&1) > v_bamtools.txt -// echo \$(plotFingerprint --version 2>&1) > v_deeptools.txt || true -// picard MarkDuplicates --version &> v_picard.txt || true -// echo \$(R --version 2>&1) > v_R.txt -// python -c "import pysam; print(pysam.__version__)" > v_pysam.txt -// echo \$(macs2 --version 2>&1) > v_macs2.txt -// touch v_homer.txt -// echo \$(ataqv --version 2>&1) > v_ataqv.txt -// echo \$(featureCounts -v 2>&1) > v_featurecounts.txt -// preseq &> v_preseq.txt -// multiqc --version > v_multiqc.txt -// scrape_software_versions.py &> software_versions_mqc.yaml -// """ -// } -// -// Channel.from(summary.collect{ [it.key, it.value] }) -// .map { k,v -> "
$k
${v ?: 'N/A'}
" } -// .reduce { a, b -> return [a, b].join("\n ") } -// .map { x -> """ -// id: 'nf-core-chipseq-summary' -// description: " - this information is collected when the pipeline is started." -// section_name: 'nf-core/chipseq Workflow Summary' -// section_href: 'https://github.com/nf-core/chipseq' -// plot_type: 'html' -// data: | -//
-// $x -//
-// """.stripIndent() } -// .set { ch_workflow_summary } -// -// /* -// * STEP 10 - MultiQC -// */ -// process MultiQC { -// publishDir "${params.outdir}/multiqc/${PEAK_TYPE}", mode: 'copy' -// -// when: -// !params.skip_multiqc -// -// input: -// file (multiqc_config) from ch_multiqc_config -// file (mqc_custom_config) from ch_multiqc_custom_config.collect().ifEmpty([]) -// -// file ('software_versions/*') from ch_software_versions_mqc.collect() -// file workflow_summary from ch_workflow_summary.collectFile(name: "workflow_summary_mqc.yaml") -// -// file ('fastqc/*') from ch_fastqc_reports_mqc.collect().ifEmpty([]) -// file ('trimgalore/*') from ch_trimgalore_results_mqc.collect().ifEmpty([]) -// file ('trimgalore/fastqc/*') from ch_trimgalore_fastqc_reports_mqc.collect().ifEmpty([]) -// -// file ('alignment/library/*') from ch_sort_bam_flagstat_mqc.collect() -// -// file ('alignment/mergedLibrary/*') from ch_mlib_bam_stats_mqc.collect() -// file ('alignment/mergedLibrary/*') from ch_mlib_rm_orphan_flagstat_mqc.collect{it[1]} -// file ('alignment/mergedLibrary/*') from ch_mlib_rm_orphan_stats_mqc.collect() -// file ('alignment/mergedLibrary/picard_metrics/*') from ch_mlib_bam_metrics_mqc.collect() -// file ('alignment/mergedLibrary/picard_metrics/*') from ch_mlib_collectmetrics_mqc.collect() -// file ('macs/mergedLibrary/*') from ch_mlib_macs_mqc.collect().ifEmpty([]) -// file ('macs/mergedLibrary/*') from ch_mlib_peak_qc_mqc.collect().ifEmpty([]) -// file ('macs/mergedLibrary/consensus/*') from ch_mlib_macs_consensus_counts_mqc.collect().ifEmpty([]) -// file ('macs/mergedLibrary/consensus/*') from ch_mlib_macs_consensus_deseq_mqc.collect().ifEmpty([]) -// file ('preseq/*') from ch_mlib_preseq_mqc.collect().ifEmpty([]) -// file ('deeptools/*') from ch_mlib_plotprofile_mqc.collect().ifEmpty([]) -// file ('deeptools/*') from ch_mlib_plotfingerprint_mqc.collect().ifEmpty([]) -// -// file ('alignment/mergedReplicate/*') from ch_mrep_bam_flagstat_mqc.collect{it[1]}.ifEmpty([]) -// file ('alignment/mergedReplicate/*') from ch_mrep_bam_stats_mqc.collect().ifEmpty([]) -// file ('alignment/mergedReplicate/*') from ch_mrep_bam_metrics_mqc.collect().ifEmpty([]) -// file ('macs/mergedReplicate/*') from ch_mrep_macs_mqc.collect().ifEmpty([]) -// file ('macs/mergedReplicate/*') from ch_mrep_peak_qc_mqc.collect().ifEmpty([]) -// file ('macs/mergedReplicate/consensus/*') from ch_mrep_macs_consensus_counts_mqc.collect().ifEmpty([]) -// file ('macs/mergedReplicate/consensus/*') from ch_mrep_macs_consensus_deseq_mqc.collect().ifEmpty([]) -// -// output: -// file "*multiqc_report.html" into ch_multiqc_report -// file "*_data" -// file "multiqc_plots" -// -// script: -// rtitle = custom_runName ? "--title \"$custom_runName\"" : '' -// rfilename = custom_runName ? "--filename " + custom_runName.replaceAll('\\W','_').replaceAll('_+','_') + "_multiqc_report" : '' -// custom_config_file = params.multiqc_config ? "--config $mqc_custom_config" : '' -// """ -// multiqc . -f $rtitle $rfilename $custom_config_file \\ -// -m custom_content -m fastqc -m cutadapt -m samtools -m picard -m preseq -m featureCounts -m deeptools -// """ -// } -// -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// /* -- -- */ -// /* -- REPORTS/DOCUMENTATION -- */ -// /* -- -- */ -// /////////////////////////////////////////////////////////////////////////////// -// /////////////////////////////////////////////////////////////////////////////// -// -// /* -// * STEP 11 - Output description HTML -// */ -// process output_documentation { -// publishDir "${params.outdir}/Documentation", mode: 'copy' -// -// input: -// file output_docs from ch_output_docs -// -// output: -// file "results_description.html" -// -// script: -// """ -// markdown_to_html.py $output_docs -o results_description.html -// """ -// } -// -// /* -// * Completion e-mail notification -// */ -// workflow.onComplete { -// -// // Set up the e-mail variables -// def subject = "[nf-core/atacseq] Successful: $workflow.runName" -// if (!workflow.success) { -// subject = "[nf-core/atacseq] FAILED: $workflow.runName" -// } -// def email_fields = [:] -// email_fields['version'] = workflow.manifest.version -// email_fields['runName'] = custom_runName ?: workflow.runName -// email_fields['success'] = workflow.success -// email_fields['dateComplete'] = workflow.complete -// email_fields['duration'] = workflow.duration -// email_fields['exitStatus'] = workflow.exitStatus -// email_fields['errorMessage'] = (workflow.errorMessage ?: 'None') -// email_fields['errorReport'] = (workflow.errorReport ?: 'None') -// email_fields['commandLine'] = workflow.commandLine -// email_fields['projectDir'] = workflow.projectDir -// email_fields['summary'] = summary -// email_fields['summary']['Date Started'] = workflow.start -// email_fields['summary']['Date Completed'] = workflow.complete -// email_fields['summary']['Pipeline script file path'] = workflow.scriptFile -// email_fields['summary']['Pipeline script hash ID'] = workflow.scriptId -// if (workflow.repository) email_fields['summary']['Pipeline repository Git URL'] = workflow.repository -// if (workflow.commitId) email_fields['summary']['Pipeline repository Git Commit'] = workflow.commitId -// if (workflow.revision) email_fields['summary']['Pipeline Git branch/tag'] = workflow.revision -// email_fields['summary']['Nextflow Version'] = workflow.nextflow.version -// email_fields['summary']['Nextflow Build'] = workflow.nextflow.build -// email_fields['summary']['Nextflow Compile Timestamp'] = workflow.nextflow.timestamp -// -// // On success try attach the multiqc report -// def mqc_report = null -// try { -// if (workflow.success) { -// mqc_report = ch_multiqc_report.getVal() -// if (mqc_report.getClass() == ArrayList) { -// log.warn "[nf-core/atacseq] Found multiple reports from process 'multiqc', will use only one" -// mqc_report = mqc_report[0] -// } -// } -// } catch (all) { -// log.warn "[nf-core/atacseq] Could not attach MultiQC report to summary email" -// } -// -// // Check if we are only sending emails on failure -// email_address = params.email -// if (!params.email && params.email_on_fail && !workflow.success) { -// email_address = params.email_on_fail -// } -// -// // Render the TXT template -// def engine = new groovy.text.GStringTemplateEngine() -// def tf = new File("$baseDir/assets/email_template.txt") -// def txt_template = engine.createTemplate(tf).make(email_fields) -// def email_txt = txt_template.toString() -// -// // Render the HTML template -// def hf = new File("$baseDir/assets/email_template.html") -// def html_template = engine.createTemplate(hf).make(email_fields) -// def email_html = html_template.toString() -// -// // Render the sendmail template -// def smail_fields = [ email: email_address, subject: subject, email_txt: email_txt, email_html: email_html, baseDir: "$baseDir", mqcFile: mqc_report, mqcMaxSize: params.max_multiqc_email_size.toBytes() ] -// def sf = new File("$baseDir/assets/sendmail_template.txt") -// def sendmail_template = engine.createTemplate(sf).make(smail_fields) -// def sendmail_html = sendmail_template.toString() -// -// // Send the HTML e-mail -// if (email_address) { -// try { -// if (params.plaintext_email) { throw GroovyException('Send plaintext e-mail, not HTML') } -// // Try to send HTML e-mail using sendmail -// [ 'sendmail', '-t' ].execute() << sendmail_html -// log.info "[nf-core/atacseq] Sent summary e-mail to $email_address (sendmail)" -// } catch (all) { -// // Catch failures and try with plaintext -// [ 'mail', '-s', subject, email_address ].execute() << email_txt -// log.info "[nf-core/atacseq] Sent summary e-mail to $email_address (mail)" -// } -// } -// -// // Write summary e-mail HTML to a file -// def output_d = new File("${params.outdir}/pipeline_info/") -// if (!output_d.exists()) { -// output_d.mkdirs() -// } -// def output_hf = new File(output_d, "pipeline_report.html") -// output_hf.withWriter { w -> w << email_html } -// def output_tf = new File(output_d, "pipeline_report.txt") -// output_tf.withWriter { w -> w << email_txt } -// -// c_green = params.monochrome_logs ? '' : "\033[0;32m"; -// c_purple = params.monochrome_logs ? '' : "\033[0;35m"; -// c_red = params.monochrome_logs ? '' : "\033[0;31m"; -// c_reset = params.monochrome_logs ? '' : "\033[0m"; -// -// if (workflow.stats.ignoredCount > 0 && workflow.success) { -// log.info "-${c_purple}Warning, pipeline completed, but with errored process(es) ${c_reset}-" -// log.info "-${c_red}Number of ignored errored process(es) : ${workflow.stats.ignoredCount} ${c_reset}-" -// log.info "-${c_green}Number of successfully ran process(es) : ${workflow.stats.succeedCount} ${c_reset}-" -// } -// if (workflow.success) { -// log.info "-${c_purple}[nf-core/atacseq]${c_green} Pipeline completed successfully${c_reset}-" -// } else { -// checkHostname() -// log.info "-${c_purple}[nf-core/atacseq]${c_red} Pipeline completed with errors${c_reset}-" -// } -// -// } +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// +/* -- -- */ +/* -- ADAPTER TRIMMING -- */ +/* -- -- */ +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// + +/* + * STEP 2 - Trim Galore! + */ +if (params.skip_trimming) { + ch_trimmed_reads = ch_raw_reads_trimgalore + ch_trimgalore_results_mqc = Channel.empty() + ch_trimgalore_fastqc_reports_mqc = Channel.empty() +} else { + process TrimGalore { + tag "$name" + label 'process_high' + publishDir "${params.outdir}/trim_galore", mode: 'copy', + saveAs: { filename -> + if (filename.endsWith(".html")) "fastqc/$filename" + else if (filename.endsWith(".zip")) "fastqc/zips/$filename" + else if (filename.endsWith("trimming_report.txt")) "logs/$filename" + else params.save_trimmed ? filename : null + } + + input: + set val(name), file(reads) from ch_raw_reads_trimgalore + + output: + set val(name), file("*.fq.gz") into ch_trimmed_reads + file "*.txt" into ch_trimgalore_results_mqc + file "*.{zip,html}" into ch_trimgalore_fastqc_reports_mqc + + script: + // Calculate number of --cores for TrimGalore based on value of task.cpus + // See: https://github.com/FelixKrueger/TrimGalore/blob/master/Changelog.md#version-060-release-on-1-mar-2019 + // See: https://github.com/nf-core/atacseq/pull/65 + def cores = 1 + if (task.cpus) { + cores = (task.cpus as int) - 4 + if (params.single_end) cores = (task.cpus as int) - 3 + if (cores < 1) cores = 1 + if (cores > 4) cores = 4 + } + + // Added soft-links to original fastqs for consistent naming in MultiQC + c_r1 = params.clip_r1 > 0 ? "--clip_r1 ${params.clip_r1}" : '' + c_r2 = params.clip_r2 > 0 ? "--clip_r2 ${params.clip_r2}" : '' + tpc_r1 = params.three_prime_clip_r1 > 0 ? "--three_prime_clip_r1 ${params.three_prime_clip_r1}" : '' + tpc_r2 = params.three_prime_clip_r2 > 0 ? "--three_prime_clip_r2 ${params.three_prime_clip_r2}" : '' + nextseq = params.trim_nextseq > 0 ? "--nextseq ${params.trim_nextseq}" : '' + + // Added soft-links to original fastqs for consistent naming in MultiQC + if (params.single_end) { + """ + [ ! -f ${name}.fastq.gz ] && ln -s $reads ${name}.fastq.gz + trim_galore --cores $cores --fastqc --gzip $c_r1 $tpc_r1 $nextseq ${name}.fastq.gz + """ + } else { + """ + [ ! -f ${name}_1.fastq.gz ] && ln -s ${reads[0]} ${name}_1.fastq.gz + [ ! -f ${name}_2.fastq.gz ] && ln -s ${reads[1]} ${name}_2.fastq.gz + trim_galore --cores $cores --paired --fastqc --gzip $c_r1 $c_r2 $tpc_r1 $tpc_r2 $nextseq ${name}_1.fastq.gz ${name}_2.fastq.gz + """ + } + } +} + +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// +/* -- -- */ +/* -- ALIGN -- */ +/* -- -- */ +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// + +/* + * STEP 3.1 - Align read 1 with bwa + */ +process BWAMem { + tag "$name" + label 'process_high' + + input: + set val(name), file(reads) from ch_trimmed_reads + file index from ch_bwa_index.collect() + + output: + set val(name), file("*.bam") into ch_bwa_bam + + script: + prefix = "${name}.Lb" + rg = "\'@RG\\tID:${name}\\tSM:${name.split('_')[0..-2].join('_')}\\tPL:ILLUMINA\\tLB:${name}\\tPU:1\'" + if (params.seq_center) { + rg = "\'@RG\\tID:${name}\\tSM:${name.split('_')[0..-2].join('_')}\\tPL:ILLUMINA\\tLB:${name}\\tPU:1\\tCN:${params.seq_center}\'" + } + """ + bwa mem \\ + -t $task.cpus \\ + -M \\ + -R $rg \\ + ${index}/${bwa_base} \\ + $reads \\ + | samtools view -@ $task.cpus -b -h -F 0x0100 -O BAM -o ${prefix}.bam - + """ +} + +/* + * STEP 3.2 - Convert .bam to coordinate sorted .bam + */ +process SortBAM { + tag "$name" + label 'process_medium' + if (params.save_align_intermeds) { + publishDir path: "${params.outdir}/bwa/library", mode: 'copy', + saveAs: { filename -> + if (filename.endsWith(".flagstat")) "samtools_stats/$filename" + else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" + else if (filename.endsWith(".stats")) "samtools_stats/$filename" + else filename + } + } + + input: + set val(name), file(bam) from ch_bwa_bam + + output: + set val(name), file("*.sorted.{bam,bam.bai}") into ch_sort_bam_merge + file "*.{flagstat,idxstats,stats}" into ch_sort_bam_flagstat_mqc + + script: + prefix = "${name}.Lb" + """ + samtools sort -@ $task.cpus -o ${prefix}.sorted.bam -T $name $bam + samtools index ${prefix}.sorted.bam + samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat + samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats + samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats + """ +} + +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// +/* -- -- */ +/* -- MERGE LIBRARY BAM -- */ +/* -- -- */ +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// + +/* + * STEP 4.1 Merge BAM files for all libraries from same replicate + */ +ch_sort_bam_merge + .map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] } + .groupTuple(by: [0]) + .map { it -> [ it[0], it[1].flatten() ] } + .set { ch_sort_bam_merge } + +process MergedLibBAM { + tag "$name" + label 'process_medium' + publishDir "${params.outdir}/bwa/mergedLibrary", mode: 'copy', + saveAs: { filename -> + if (filename.endsWith(".flagstat")) "samtools_stats/$filename" + else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" + else if (filename.endsWith(".stats")) "samtools_stats/$filename" + else if (filename.endsWith(".metrics.txt")) "picard_metrics/$filename" + else params.save_align_intermeds ? filename : null + } + + input: + set val(name), file(bams) from ch_sort_bam_merge + + output: + set val(name), file("*${prefix}.sorted.{bam,bam.bai}") into ch_mlib_bam_filter, + ch_mlib_bam_preseq, + ch_mlib_bam_ataqv + file "*.{flagstat,idxstats,stats}" into ch_mlib_bam_stats_mqc + file "*.txt" into ch_mlib_bam_metrics_mqc + + script: + prefix = "${name}.mLb.mkD" + bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() + def avail_mem = 3 + if (!task.memory) { + log.info "[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." + } else { + avail_mem = task.memory.toGiga() + } + if (bam_files.size() > 1) { + """ + picard -Xmx${avail_mem}g MergeSamFiles \\ + ${'INPUT='+bam_files.join(' INPUT=')} \\ + OUTPUT=${name}.sorted.bam \\ + SORT_ORDER=coordinate \\ + VALIDATION_STRINGENCY=LENIENT \\ + TMP_DIR=tmp + samtools index ${name}.sorted.bam + + picard -Xmx${avail_mem}g MarkDuplicates \\ + INPUT=${name}.sorted.bam \\ + OUTPUT=${prefix}.sorted.bam \\ + ASSUME_SORTED=true \\ + REMOVE_DUPLICATES=false \\ + METRICS_FILE=${prefix}.MarkDuplicates.metrics.txt \\ + VALIDATION_STRINGENCY=LENIENT \\ + TMP_DIR=tmp + + samtools index ${prefix}.sorted.bam + samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats + samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat + samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats + """ + } else { + """ + picard -Xmx${avail_mem}g MarkDuplicates \\ + INPUT=${bam_files[0]} \\ + OUTPUT=${prefix}.sorted.bam \\ + ASSUME_SORTED=true \\ + REMOVE_DUPLICATES=false \\ + METRICS_FILE=${prefix}.MarkDuplicates.metrics.txt \\ + VALIDATION_STRINGENCY=LENIENT \\ + TMP_DIR=tmp + + samtools index ${prefix}.sorted.bam + samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats + samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat + samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats + """ + } +} + +/* + * STEP 4.2 Filter BAM file at merged library-level + */ +process MergedLibBAMFilter { + tag "$name" + label 'process_medium' + publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', + saveAs: { filename -> + if (params.single_end || params.save_align_intermeds) { + if (filename.endsWith(".flagstat")) "samtools_stats/$filename" + else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" + else if (filename.endsWith(".stats")) "samtools_stats/$filename" + else if (filename.endsWith(".sorted.bam")) filename + else if (filename.endsWith(".sorted.bam.bai")) filename + else null + } + } + + input: + set val(name), file(bam) from ch_mlib_bam_filter + file bed from ch_genome_filter_regions.collect() + file bamtools_filter_config from ch_bamtools_filter_config + + output: + set val(name), file("*.{bam,bam.bai}") into ch_mlib_filter_bam + set val(name), file("*.flagstat") into ch_mlib_filter_bam_flagstat + file "*.{idxstats,stats}" into ch_mlib_filter_bam_stats_mqc + + script: + prefix = params.single_end ? "${name}.mLb.clN" : "${name}.mLb.flT" + filter_params = params.single_end ? "-F 0x004" : "-F 0x004 -F 0x0008 -f 0x001" + dup_params = params.keep_dups ? "" : "-F 0x0400" + multimap_params = params.keep_multi_map ? "" : "-q 1" + blacklist_params = params.blacklist ? "-L $bed" : "" + name_sort_bam = params.single_end ? "" : "samtools sort -n -@ $task.cpus -o ${prefix}.bam -T $prefix ${prefix}.sorted.bam" + """ + samtools view \\ + $filter_params \\ + $dup_params \\ + $multimap_params \\ + $blacklist_params \\ + -b ${bam[0]} \\ + | bamtools filter \\ + -out ${prefix}.sorted.bam \\ + -script $bamtools_filter_config + + samtools index ${prefix}.sorted.bam + samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat + samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats + samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats + + $name_sort_bam + """ +} + +/* + * STEP 4.3 Remove orphan reads from paired-end BAM file + */ +if (params.single_end) { + ch_mlib_filter_bam + .into { ch_mlib_rm_orphan_bam_metrics; + ch_mlib_rm_orphan_bam_bigwig; + ch_mlib_rm_orphan_bam_macs; + ch_mlib_rm_orphan_bam_plotfingerprint; + ch_mlib_rm_orphan_bam_mrep; + ch_mlib_name_bam_mlib_counts; + ch_mlib_name_bam_mrep_counts } + + ch_mlib_filter_bam_flagstat + .into { ch_mlib_rm_orphan_flagstat_bigwig; + ch_mlib_rm_orphan_flagstat_macs; + ch_mlib_rm_orphan_flagstat_mqc } + + ch_mlib_filter_bam_stats_mqc + .set { ch_mlib_rm_orphan_stats_mqc } +} else { + process MergedLibBAMRemoveOrphan { + tag "$name" + label 'process_medium' + publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', + saveAs: { filename -> + if (filename.endsWith(".flagstat")) "samtools_stats/$filename" + else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" + else if (filename.endsWith(".stats")) "samtools_stats/$filename" + else if (filename.endsWith(".sorted.bam")) filename + else if (filename.endsWith(".sorted.bam.bai")) filename + else null + } + + input: + set val(name), file(bam) from ch_mlib_filter_bam + + output: + set val(name), file("*.sorted.{bam,bam.bai}") into ch_mlib_rm_orphan_bam_metrics, + ch_mlib_rm_orphan_bam_bigwig, + ch_mlib_rm_orphan_bam_macs, + ch_mlib_rm_orphan_bam_plotfingerprint, + ch_mlib_rm_orphan_bam_mrep + set val(name), file("${prefix}.bam") into ch_mlib_name_bam_mlib_counts, + ch_mlib_name_bam_mrep_counts + set val(name), file("*.flagstat") into ch_mlib_rm_orphan_flagstat_bigwig, + ch_mlib_rm_orphan_flagstat_macs, + ch_mlib_rm_orphan_flagstat_mqc + file "*.{idxstats,stats}" into ch_mlib_rm_orphan_stats_mqc + + script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ + prefix = "${name}.mLb.clN" + """ + bampe_rm_orphan.py ${bam[0]} ${prefix}.bam --only_fr_pairs + + samtools sort -@ $task.cpus -o ${prefix}.sorted.bam -T $prefix ${prefix}.bam + samtools index ${prefix}.sorted.bam + samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat + samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats + samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats + """ + } +} + +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// +/* -- -- */ +/* -- MERGE LIBRARY BAM POST-ANALYSIS -- */ +/* -- -- */ +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// + +/* + * STEP 5.1 preseq analysis after merging libraries and before filtering + */ +process MergedLibPreseq { + tag "$name" + label 'process_low' + publishDir "${params.outdir}/bwa/mergedLibrary/preseq", mode: 'copy' + + when: + !params.skip_preseq + + input: + set val(name), file(bam) from ch_mlib_bam_preseq + + output: + file "*.ccurve.txt" into ch_mlib_preseq_mqc + + script: + prefix = "${name}.mLb.mkD" + """ + preseq lc_extrap -v -output ${prefix}.ccurve.txt -bam ${bam[0]} + """ +} + +/* + * STEP 5.2 Picard CollectMultipleMetrics after merging libraries and filtering + */ +process MergedLibMetrics { + tag "$name" + label 'process_medium' + publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', + saveAs: { filename -> + if (filename.endsWith("_metrics")) "picard_metrics/$filename" + else if (filename.endsWith(".pdf")) "picard_metrics/pdf/$filename" + else null + } + + when: + !params.skip_picard_metrics + + input: + set val(name), file(bam) from ch_mlib_rm_orphan_bam_metrics + file fasta from ch_fasta + + output: + file "*_metrics" into ch_mlib_collectmetrics_mqc + file "*.pdf" into ch_mlib_collectmetrics_pdf + + script: + prefix = "${name}.mLb.clN" + def avail_mem = 3 + if (!task.memory) { + log.info "[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." + } else { + avail_mem = task.memory.toGiga() + } + """ + picard -Xmx${avail_mem}g CollectMultipleMetrics \\ + INPUT=${bam[0]} \\ + OUTPUT=${prefix}.CollectMultipleMetrics \\ + REFERENCE_SEQUENCE=$fasta \\ + VALIDATION_STRINGENCY=LENIENT \\ + TMP_DIR=tmp + """ +} + +/* + * STEP 5.3 Read depth normalised bigWig + */ +process MergedLibBigWig { + tag "$name" + label 'process_medium' + publishDir "${params.outdir}/bwa/mergedLibrary/bigwig", mode: 'copy', + saveAs: { filename -> + if (filename.endsWith("scale_factor.txt")) "scale/$filename" + else if (filename.endsWith(".bigWig")) "$filename" + else null + } + + input: + set val(name), file(bam), file(flagstat) from ch_mlib_rm_orphan_bam_bigwig.join(ch_mlib_rm_orphan_flagstat_bigwig, by: [0]) + file sizes from ch_genome_sizes_mlib_bigwig.collect() + + output: + set val(name), file("*.bigWig") into ch_mlib_bigwig_plotprofile + file "*scale_factor.txt" into ch_mlib_bigwig_scale + file "*igv.txt" into ch_mlib_bigwig_igv + + script: + prefix = "${name}.mLb.clN" + pe_fragment = params.single_end ? "" : "-pc" + extend = (params.single_end && params.fragment_size > 0) ? "-fs ${params.fragment_size}" : '' + """ + SCALE_FACTOR=\$(grep 'mapped (' $flagstat | awk '{print 1000000/\$1}') + echo \$SCALE_FACTOR > ${prefix}.scale_factor.txt + genomeCoverageBed -ibam ${bam[0]} -bg -scale \$SCALE_FACTOR $pe_fragment $extend | sort -T '.' -k1,1 -k2,2n > ${prefix}.bedGraph + + bedGraphToBigWig ${prefix}.bedGraph $sizes ${prefix}.bigWig + + find * -type f -name "*.bigWig" -exec echo -e "bwa/mergedLibrary/bigwig/"{}"\\t0,0,178" \\; > ${prefix}.bigWig.igv.txt + """ +} + +/* + * STEP 5.4 generate gene body coverage plot with deepTools + */ +process MergedLibPlotProfile { + tag "$name" + label 'process_high' + publishDir "${params.outdir}/bwa/mergedLibrary/deepTools/plotProfile", mode: 'copy' + + when: + !params.skip_plot_profile + + input: + set val(name), file(bigwig) from ch_mlib_bigwig_plotprofile + file bed from ch_gene_bed + + output: + file '*.{gz,pdf}' into ch_mlib_plotprofile_results + file '*.plotProfile.tab' into ch_mlib_plotprofile_mqc + + script: + prefix = "${name}.mLb.clN" + """ + computeMatrix scale-regions \\ + --regionsFileName $bed \\ + --scoreFileName $bigwig \\ + --outFileName ${prefix}.computeMatrix.mat.gz \\ + --outFileNameMatrix ${prefix}.computeMatrix.vals.mat.gz \\ + --regionBodyLength 1000 \\ + --beforeRegionStartLength 3000 \\ + --afterRegionStartLength 3000 \\ + --skipZeros \\ + --smartLabels \\ + --numberOfProcessors $task.cpus + + plotProfile --matrixFile ${prefix}.computeMatrix.mat.gz \\ + --outFileName ${prefix}.plotProfile.pdf \\ + --outFileNameData ${prefix}.plotProfile.tab + """ +} + +/* + * STEP 5.5 deepTools plotFingerprint + */ +process MergedLibPlotFingerprint { + tag "$name" + label 'process_high' + publishDir "${params.outdir}/bwa/mergedLibrary/deepTools/plotFingerprint", mode: 'copy' + + when: + !params.skip_plot_fingerprint + + input: + set val(name), file(bam) from ch_mlib_rm_orphan_bam_plotfingerprint + + output: + file '*.{txt,pdf}' into ch_mlib_plotfingerprint_results + file '*.raw.txt' into ch_mlib_plotfingerprint_mqc + + script: + prefix = "${name}.mLb.clN" + extend = (params.single_end && params.fragment_size > 0) ? "--extendReads ${params.fragment_size}" : '' + """ + plotFingerprint \\ + --bamfiles ${bam[0]} \\ + --plotFile ${prefix}.plotFingerprint.pdf \\ + $extend \\ + --labels $prefix \\ + --outRawCounts ${prefix}.plotFingerprint.raw.txt \\ + --outQualityMetrics ${prefix}.plotFingerprint.qcmetrics.txt \\ + --skipZeros \\ + --numberOfProcessors $task.cpus \\ + --numberOfSamples $params.fingerprint_bins + """ +} + +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// +/* -- -- */ +/* -- MERGE LIBRARY PEAK ANALYSIS -- */ +/* -- -- */ +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// + +/* + * STEP 6.1 Call peaks with MACS2 and calculate FRiP score + */ +process MergedLibMACSCallPeak { + tag "$name" + label 'process_medium' + publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}", mode: 'copy', + saveAs: { filename -> + if (filename.endsWith(".tsv")) "qc/$filename" + else if (filename.endsWith(".igv.txt")) null + else filename + } + + when: + params.macs_gsize + + input: + set val(name), file(bam), file(flagstat) from ch_mlib_rm_orphan_bam_macs.join(ch_mlib_rm_orphan_flagstat_macs, by: [0]) + file mlib_peak_count_header from ch_mlib_peak_count_header + file mlib_frip_score_header from ch_mlib_frip_score_header + + output: + set val(name), file("*.{bed,xls,gappedPeak,bdg}") into ch_mlib_macs_output + set val(name), file("*$PEAK_TYPE") into ch_mlib_macs_homer, + ch_mlib_macs_qc, + ch_mlib_macs_consensus, + ch_mlib_macs_ataqv + file "*igv.txt" into ch_mlib_macs_igv + file "*_mqc.tsv" into ch_mlib_macs_mqc + + script: + prefix = "${name}.mLb.clN" + broad = params.narrow_peak ? '' : "--broad --broad-cutoff ${params.broad_cutoff}" + format = params.single_end ? "BAM" : "BAMPE" + pileup = params.save_macs_pileup ? "-B --SPMR" : "" + """ + macs2 callpeak \\ + -t ${bam[0]} \\ + $broad \\ + -f $format \\ + -g $params.macs_gsize \\ + -n $prefix \\ + $pileup \\ + --keep-dup all \\ + --nomodel + + cat ${prefix}_peaks.${PEAK_TYPE} | wc -l | awk -v OFS='\t' '{ print "${name}", \$1 }' | cat $mlib_peak_count_header - > ${prefix}_peaks.count_mqc.tsv + + READS_IN_PEAKS=\$(intersectBed -a ${bam[0]} -b ${prefix}_peaks.${PEAK_TYPE} -bed -c -f 0.20 | awk -F '\t' '{sum += \$NF} END {print sum}') + grep 'mapped (' $flagstat | awk -v a="\$READS_IN_PEAKS" -v OFS='\t' '{print "${name}", a/\$1}' | cat $mlib_frip_score_header - > ${prefix}_peaks.FRiP_mqc.tsv + + find * -type f -name "*.${PEAK_TYPE}" -exec echo -e "bwa/mergedLibrary/macs/${PEAK_TYPE}/"{}"\\t0,0,178" \\; > ${prefix}_peaks.igv.txt + """ +} + +/* + * STEP 6.2 Annotate peaks with HOMER + */ +process MergedLibAnnotatePeaks { + tag "$name" + label "process_medium" + publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}", mode: 'copy' + + when: + params.macs_gsize + + input: + set val(name), file(peak) from ch_mlib_macs_homer + file fasta from ch_fasta + file gtf from ch_gtf + + output: + file "*.txt" into ch_mlib_macs_annotate + + script: + prefix = "${name}.mLb.clN" + """ + annotatePeaks.pl \\ + $peak \\ + $fasta \\ + -gid \\ + -gtf $gtf \\ + -cpu $task.cpus \\ + > ${prefix}_peaks.annotatePeaks.txt + """ +} + +/* + * STEP 6.3 Aggregated QC plots for peaks, FRiP and peak-to-gene annotation + */ +process MergedLibPeakQC { + label "process_medium" + publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/qc", mode: 'copy' + + when: + params.macs_gsize + + input: + file peaks from ch_mlib_macs_qc.collect{ it[1] } + file annos from ch_mlib_macs_annotate.collect() + file mlib_peak_annotation_header from ch_mlib_peak_annotation_header + + output: + file "*.{txt,pdf}" into ch_mlib_peak_qc + file "*.tsv" into ch_mlib_peak_qc_mqc + + script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ + suffix = 'mLb.clN' + """ + plot_macs_qc.r \\ + -i ${peaks.join(',')} \\ + -s ${peaks.join(',').replaceAll(".${suffix}_peaks.${PEAK_TYPE}","")} \\ + -o ./ \\ + -p macs_peak.${suffix} + + plot_homer_annotatepeaks.r \\ + -i ${annos.join(',')} \\ + -s ${annos.join(',').replaceAll(".${suffix}_peaks.annotatePeaks.txt","")} \\ + -o ./ \\ + -p macs_annotatePeaks.${suffix} + + cat $mlib_peak_annotation_header macs_annotatePeaks.${suffix}.summary.txt > macs_annotatePeaks.${suffix}.summary_mqc.tsv + """ +} + +/* + * STEP 6.4 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection + */ +process MergedLibConsensusPeakSet { + label 'process_long' + publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: 'copy', + saveAs: { filename -> + if (filename.endsWith(".igv.txt")) null + else filename + } + + when: + params.macs_gsize && (replicatesExist || multipleGroups) && !params.skip_consensus_peaks + + input: + file peaks from ch_mlib_macs_consensus.collect{ it[1] } + + output: + file "*.bed" into ch_mlib_macs_consensus_bed + file "*.saf" into ch_mlib_macs_consensus_saf + file "*.boolean.txt" into ch_mlib_macs_consensus_bool + file "*.intersect.{txt,plot.pdf}" into ch_mlib_macs_consensus_intersect + file "*igv.txt" into ch_mlib_macs_consensus_igv + + script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ + suffix = 'mLb.clN' + prefix = "consensus_peaks.${suffix}" + mergecols = params.narrow_peak ? (2..10).join(',') : (2..9).join(',') + collapsecols = params.narrow_peak ? (["collapse"]*9).join(',') : (["collapse"]*8).join(',') + expandparam = params.narrow_peak ? "--is_narrow_peak" : "" + """ + sort -T '.' -k1,1 -k2,2n ${peaks.collect{it.toString()}.sort().join(' ')} \\ + | mergeBed -c $mergecols -o $collapsecols > ${prefix}.txt + + macs2_merged_expand.py \\ + ${prefix}.txt \\ + ${peaks.collect{it.toString()}.sort().join(',').replaceAll("_peaks.${PEAK_TYPE}","")} \\ + ${prefix}.boolean.txt \\ + --min_replicates $params.min_reps_consensus \\ + $expandparam + + awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$1, \$2, \$3, \$4, "0", "+" }' ${prefix}.boolean.txt > ${prefix}.bed + + echo -e "GeneID\tChr\tStart\tEnd\tStrand" > ${prefix}.saf + awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$4, \$1, \$2, \$3, "+" }' ${prefix}.boolean.txt >> ${prefix}.saf + + sed -i 's/.${suffix}//g' ${prefix}.boolean.intersect.txt + plot_peak_intersect.r -i ${prefix}.boolean.intersect.txt -o ${prefix}.boolean.intersect.plot.pdf + + find * -type f -name "${prefix}.bed" -exec echo -e "bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus/"{}"\\t0,0,0" \\; > ${prefix}.bed.igv.txt + """ +} + +/* + * STEP 6.5 Annotate consensus peaks with HOMER, and add annotation to boolean output file + */ +process MergedLibConsensusPeakSetAnnotate { + label "process_medium" + publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: 'copy' + + when: + params.macs_gsize && (replicatesExist || multipleGroups) && !params.skip_consensus_peaks + + input: + file bed from ch_mlib_macs_consensus_bed + file bool from ch_mlib_macs_consensus_bool + file fasta from ch_fasta + file gtf from ch_gtf + + output: + file "*.annotatePeaks.txt" into ch_mlib_macs_consensus_annotate + + script: + prefix = "consensus_peaks.mLb.clN" + """ + annotatePeaks.pl \\ + $bed \\ + $fasta \\ + -gid \\ + -gtf $gtf \\ + -cpu $task.cpus \\ + > ${prefix}.annotatePeaks.txt + + cut -f2- ${prefix}.annotatePeaks.txt | awk 'NR==1; NR > 1 {print \$0 | "sort -T '.' -k1,1 -k2,2n"}' | cut -f6- > tmp.txt + paste $bool tmp.txt > ${prefix}.boolean.annotatePeaks.txt + """ +} + +/* + * STEP 6.6 Count reads in consensus peaks with featureCounts + */ +process MergedLibConsensusPeakSetCounts { + label 'process_medium' + publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: 'copy' + + when: + params.macs_gsize && (replicatesExist || multipleGroups) && !params.skip_consensus_peaks + + input: + file bams from ch_mlib_name_bam_mlib_counts.collect{ it[1] } + file saf from ch_mlib_macs_consensus_saf.collect() + + output: + file "*featureCounts.txt" into ch_mlib_macs_consensus_counts + file "*featureCounts.txt.summary" into ch_mlib_macs_consensus_counts_mqc + + script: + prefix = "consensus_peaks.mLb.clN" + bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() + pe_params = params.single_end ? '' : "-p --donotsort" + """ + featureCounts \\ + -F SAF \\ + -O \\ + --fracOverlap 0.2 \\ + -T $task.cpus \\ + $pe_params \\ + -a $saf \\ + -o ${prefix}.featureCounts.txt \\ + ${bam_files.join(' ')} + """ +} + +/* + * STEP 6.7 Perform differential analysis with DESeq2 + */ +process MergedLibConsensusPeakSetDESeq { + label 'process_medium' + publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus/deseq2", mode: 'copy', + saveAs: { filename -> + if (filename.endsWith(".igv.txt")) null + else filename + } + + when: + params.macs_gsize && replicatesExist && multipleGroups && !params.skip_consensus_peaks && !params.skip_diff_analysis + + input: + file counts from ch_mlib_macs_consensus_counts + file mlib_deseq2_pca_header from ch_mlib_deseq2_pca_header + file mlib_deseq2_clustering_header from ch_mlib_deseq2_clustering_header + + output: + file "*.{RData,results.txt,pdf,log}" into ch_mlib_macs_consensus_deseq_results + file "sizeFactors" into ch_mlib_macs_consensus_deseq_factors + file "*vs*/*.{pdf,txt}" into ch_mlib_macs_consensus_deseq_comp_results + file "*vs*/*.bed" into ch_mlib_macs_consensus_deseq_comp_bed + file "*igv.txt" into ch_mlib_macs_consensus_deseq_comp_igv + file "*.tsv" into ch_mlib_macs_consensus_deseq_mqc + + script: + prefix = "consensus_peaks.mLb.clN" + bam_ext = params.single_end ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" + """ + featurecounts_deseq2.r \\ + --featurecount_file $counts \\ + --bam_suffix '$bam_ext' \\ + --outdir ./ \\ + --outprefix $prefix \\ + --outsuffix .mLb \\ + --cores $task.cpus + + cat $mlib_deseq2_pca_header ${prefix}.pca.vals.txt > ${prefix}.pca.vals_mqc.tsv + cat $mlib_deseq2_clustering_header ${prefix}.sample.dists.txt > ${prefix}.sample.dists_mqc.tsv + + find * -type f -name "*.FDR0.05.results.bed" -exec echo -e "bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus/deseq2/"{}"\\t255,0,0" \\; > ${prefix}.igv.txt + """ +} + +/* + * STEP 6.8 Run ataqv on BAM file and corresponding peaks + */ +process MergedLibAtaqv { + tag "$name" + label 'process_medium' + publishDir "${params.outdir}/bwa/mergedLibrary/ataqv/${PEAK_TYPE}", mode: 'copy' + + when: + !params.skip_ataqv + + input: + set val(name), file(bam), file(peak) from ch_mlib_bam_ataqv.join(ch_mlib_macs_ataqv, by: [0]) + file autosomes from ch_genome_autosomes.collect() + file tss_bed from ch_tss_bed + + output: + file "*.json" into ch_mlib_ataqv + + script: + peak_param = params.macs_gsize ? "--peak-file ${peak}" : "" + mito_param = params.mito_name ? "--mitochondrial-reference-name ${params.mito_name}" : "" + """ + ataqv \\ + --threads $task.cpus \\ + $peak_param \\ + --tss-file $tss_bed \\ + --metrics-file ${name}.ataqv.json \\ + --name $name \\ + --ignore-read-groups \\ + --autosomal-reference-file $autosomes \\ + $mito_param \\ + NA \\ + ${bam[0]} + """ +} + +/* + * STEP 6.9 run ataqv mkarv on all JSON files to render web app + */ +process MergedLibAtaqvMkarv { + label 'process_medium' + publishDir "${params.outdir}/bwa/mergedLibrary/ataqv/${PEAK_TYPE}", mode: 'copy' + + when: + !params.skip_ataqv + + input: + file json from ch_mlib_ataqv.collect() + + output: + file "html" into ch_mlib_ataqv_mkarv + + script: + """ + mkarv \\ + --concurrency $task.cpus \\ + --force \\ + ./html/ \\ + ${json.join(' ')} + """ +} + +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// +/* -- -- */ +/* -- MERGE REPLICATE BAM -- */ +/* -- -- */ +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// + +/* + * STEP 7 Merge library BAM files across all replicates + */ +ch_mlib_rm_orphan_bam_mrep + .map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] } + .groupTuple(by: [0]) + .map { it -> [ it[0], it[1].flatten() ] } + .set { ch_mlib_rm_orphan_bam_mrep } + +process MergedRepBAM { + tag "$name" + label 'process_medium' + publishDir "${params.outdir}/bwa/mergedReplicate", mode: 'copy', + saveAs: { filename -> + if (filename.endsWith(".flagstat")) "samtools_stats/$filename" + else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" + else if (filename.endsWith(".stats")) "samtools_stats/$filename" + else if (filename.endsWith(".metrics.txt")) "picard_metrics/$filename" + else filename + } + + input: + set val(name), file(bams) from ch_mlib_rm_orphan_bam_mrep + + output: + set val(name), file("*${prefix}.sorted.{bam,bam.bai}") into ch_mrep_bam_bigwig, + ch_mrep_bam_macs + set val(name), file("*.flagstat") into ch_mrep_bam_flagstat_bigwig, + ch_mrep_bam_flagstat_macs, + ch_mrep_bam_flagstat_mqc + file "*.{idxstats,stats}" into ch_mrep_bam_stats_mqc + file "*.txt" into ch_mrep_bam_metrics_mqc + + when: + !params.skip_merge_replicates && replicatesExist + + script: + prefix = "${name}.mRp.clN" + bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() + def avail_mem = 3 + if (!task.memory) { + log.info "[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." + } else { + avail_mem = task.memory.toGiga() + } + if (bam_files.size() > 1) { + """ + picard -Xmx${avail_mem}g MergeSamFiles \\ + ${'INPUT='+bam_files.join(' INPUT=')} \\ + OUTPUT=${name}.sorted.bam \\ + SORT_ORDER=coordinate \\ + VALIDATION_STRINGENCY=LENIENT \\ + TMP_DIR=tmp + samtools index ${name}.sorted.bam + + picard -Xmx${avail_mem}g MarkDuplicates \\ + INPUT=${name}.sorted.bam \\ + OUTPUT=${prefix}.sorted.bam \\ + ASSUME_SORTED=true \\ + REMOVE_DUPLICATES=true \\ + METRICS_FILE=${prefix}.MarkDuplicates.metrics.txt \\ + VALIDATION_STRINGENCY=LENIENT \\ + TMP_DIR=tmp + + samtools index ${prefix}.sorted.bam + samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat + samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats + samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats + """ + } else { + """ + ln -s ${bams[0]} ${prefix}.sorted.bam + ln -s ${bams[1]} ${prefix}.sorted.bam.bai + touch ${prefix}.MarkDuplicates.metrics.txt + samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat + samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats + samtools stats ${prefix}.sorted.bam > ${prefix}.sorted.bam.stats + """ + } +} + +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// +/* -- -- */ +/* -- MERGE REPLICATE BAM POST-ANALYSIS -- */ +/* -- -- */ +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// + +/* + * STEP 8.1 Read depth normalised bigWig + */ +process MergedRepBigWig { + tag "$name" + label 'process_medium' + publishDir "${params.outdir}/bwa/mergedReplicate/bigwig", mode: 'copy', + saveAs: { filename -> + if (filename.endsWith("scale_factor.txt")) "scale/$filename" + else if (filename.endsWith(".bigWig")) "$filename" + else null + } + + when: + !params.skip_merge_replicates && replicatesExist + + input: + set val(name), file(bam), file(flagstat) from ch_mrep_bam_bigwig.join(ch_mrep_bam_flagstat_bigwig, by: [0]) + file sizes from ch_genome_sizes_mrep_bigwig.collect() + + output: + set val(name), file("*.bigWig") into ch_mrep_bigwig + file "*scale_factor.txt" into ch_mrep_bigwig_scale + file "*igv.txt" into ch_mrep_bigwig_igv + + script: + prefix = "${name}.mRp.clN" + pe_fragment = params.single_end ? "" : "-pc" + extend = (params.single_end && params.fragment_size > 0) ? "-fs ${params.fragment_size}" : '' + """ + SCALE_FACTOR=\$(grep 'mapped (' $flagstat | awk '{print 1000000/\$1}') + echo \$SCALE_FACTOR > ${prefix}.scale_factor.txt + genomeCoverageBed -ibam ${bam[0]} -bg -scale \$SCALE_FACTOR $pe_fragment $extend | sort -T '.' -k1,1 -k2,2n > ${prefix}.bedGraph + + bedGraphToBigWig ${prefix}.bedGraph $sizes ${prefix}.bigWig + + find * -type f -name "*.bigWig" -exec echo -e "bwa/mergedReplicate/bigwig/"{}"\\t0,0,178" \\; > ${prefix}.bigWig.igv.txt + """ +} + +/* + * STEP 8.2 Call peaks with MACS2 and calculate FRiP score + */ +process MergedRepMACSCallPeak { + tag "$name" + label 'process_medium' + publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}", mode: 'copy', + saveAs: { filename -> + if (filename.endsWith(".tsv")) "qc/$filename" + else if (filename.endsWith(".igv.txt")) null + else filename + } + + when: + !params.skip_merge_replicates && replicatesExist && params.macs_gsize + + input: + set val(name), file(bam), file(flagstat) from ch_mrep_bam_macs.join(ch_mrep_bam_flagstat_macs, by: [0]) + file mrep_peak_count_header from ch_mrep_peak_count_header + file mrep_frip_score_header from ch_mrep_frip_score_header + + output: + set val(name), file("*.{bed,xls,gappedPeak,bdg}") into ch_mrep_macs_output + set val(name), file("*$PEAK_TYPE") into ch_mrep_macs_homer, + ch_mrep_macs_qc, + ch_mrep_macs_consensus + file "*igv.txt" into ch_mrep_macs_igv + file "*_mqc.tsv" into ch_mrep_macs_mqc + + script: + prefix = "${name}.mRp.clN" + broad = params.narrow_peak ? '' : "--broad --broad-cutoff ${params.broad_cutoff}" + format = params.single_end ? "BAM" : "BAMPE" + pileup = params.save_macs_pileup ? "-B --SPMR" : "" + """ + macs2 callpeak \\ + -t ${bam[0]} \\ + $broad \\ + -f $format \\ + -g $params.macs_gsize \\ + -n $prefix \\ + $pileup \\ + --keep-dup all \\ + --nomodel + + cat ${prefix}_peaks.${PEAK_TYPE} | wc -l | awk -v OFS='\t' '{ print "${name}", \$1 }' | cat $mrep_peak_count_header - > ${prefix}_peaks.count_mqc.tsv + + READS_IN_PEAKS=\$(intersectBed -a ${bam[0]} -b ${prefix}_peaks.${PEAK_TYPE} -bed -c -f 0.20 | awk -F '\t' '{sum += \$NF} END {print sum}') + grep 'mapped (' $flagstat | awk -v a="\$READS_IN_PEAKS" -v OFS='\t' '{print "${name}", a/\$1}' | cat $mrep_frip_score_header - > ${prefix}_peaks.FRiP_mqc.tsv + + find * -type f -name "*.${PEAK_TYPE}" -exec echo -e "bwa/mergedReplicate/macs/${PEAK_TYPE}/"{}"\\t0,0,178" \\; > ${prefix}_peaks.igv.txt + """ +} + +/* + * STEP 8.3 Annotate peaks with HOMER + */ +process MergedRepAnnotatePeaks { + tag "$name" + label "process_medium" + publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}", mode: 'copy' + + when: + !params.skip_merge_replicates && replicatesExist && params.macs_gsize + + input: + set val(name), file(peak) from ch_mrep_macs_homer + file fasta from ch_fasta + file gtf from ch_gtf + + output: + file "*.txt" into ch_mrep_macs_annotate + + script: + prefix = "${name}.mRp.clN" + """ + annotatePeaks.pl \\ + $peak \\ + $fasta \\ + -gid \\ + -gtf $gtf \\ + -cpu $task.cpus \\ + > ${prefix}_peaks.annotatePeaks.txt + """ +} + +/* + * STEP 8.4 Aggregated QC plots for peaks, FRiP and peak-to-gene annotation + */ +process MergedRepPeakQC { + label "process_medium" + publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/qc", mode: 'copy' + + when: + !params.skip_merge_replicates && replicatesExist && params.macs_gsize + + input: + file peaks from ch_mrep_macs_qc.collect{ it[1] } + file annos from ch_mrep_macs_annotate.collect() + file mrep_peak_annotation_header from ch_mrep_peak_annotation_header + + output: + file "*.{txt,pdf}" into ch_mrep_peak_qc + file "*.tsv" into ch_mrep_peak_qc_mqc + + script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ + suffix = 'mRp.clN' + """ + plot_macs_qc.r \\ + -i ${peaks.join(',')} \\ + -s ${peaks.join(',').replaceAll(".${suffix}_peaks.${PEAK_TYPE}","")} \\ + -o ./ \\ + -p macs_peak.${suffix} + + plot_homer_annotatepeaks.r \\ + -i ${annos.join(',')} \\ + -s ${annos.join(',').replaceAll(".${suffix}_peaks.annotatePeaks.txt","")} \\ + -o ./ \\ + -p macs_annotatePeaks.${suffix} + + cat $mrep_peak_annotation_header macs_annotatePeaks.${suffix}.summary.txt > macs_annotatePeaks.${suffix}.summary_mqc.tsv + """ +} + +/* + * STEP 8.5 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection + */ +process MergedRepConsensusPeakSet { + label 'process_long' + publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: 'copy', + saveAs: { filename -> + if (filename.endsWith(".igv.txt")) null + else filename + } + + when: + !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks + + input: + file peaks from ch_mrep_macs_consensus.collect{ it[1] } + + output: + file "*.bed" into ch_mrep_macs_consensus_bed + file "*.saf" into ch_mrep_macs_consensus_saf + file "*.boolean.txt" into ch_mrep_macs_consensus_bool + file "*.intersect.{txt,plot.pdf}" into ch_mrep_macs_consensus_intersect + file "*igv.txt" into ch_mrep_macs_consensus_igv + + script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ + suffix = 'mRp.clN' + prefix = "consensus_peaks.${suffix}" + mergecols = params.narrow_peak ? (2..10).join(',') : (2..9).join(',') + collapsecols = params.narrow_peak ? (["collapse"]*9).join(',') : (["collapse"]*8).join(',') + expandparam = params.narrow_peak ? "--is_narrow_peak" : "" + """ + sort -T '.' -k1,1 -k2,2n ${peaks.collect{it.toString()}.sort().join(' ')} \\ + | mergeBed -c $mergecols -o $collapsecols > ${prefix}.txt + + macs2_merged_expand.py \\ + ${prefix}.txt \\ + ${peaks.collect{it.toString()}.sort().join(',').replaceAll("_peaks.${PEAK_TYPE}","")} \\ + ${prefix}.boolean.txt \\ + $expandparam + + awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$1, \$2, \$3, \$4, "0", "+" }' ${prefix}.boolean.txt > ${prefix}.bed + + echo -e "GeneID\tChr\tStart\tEnd\tStrand" > ${prefix}.saf + awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$4, \$1, \$2, \$3, "+" }' ${prefix}.boolean.txt >> ${prefix}.saf + + sed -i 's/.${suffix}//g' ${prefix}.boolean.intersect.txt + plot_peak_intersect.r -i ${prefix}.boolean.intersect.txt -o ${prefix}.boolean.intersect.plot.pdf + + find * -type f -name "${prefix}.bed" -exec echo -e "bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus/"{}"\\t0,0,0" \\; > ${prefix}.bed.igv.txt + """ +} + +/* + * STEP 8.6 Annotate consensus peaks with HOMER, and add annotation to boolean output file + */ +process MergedRepConsensusPeakSetAnnotate { + label "process_medium" + publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: 'copy' + + when: + !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks + + input: + file bed from ch_mrep_macs_consensus_bed + file bool from ch_mrep_macs_consensus_bool + file fasta from ch_fasta + file gtf from ch_gtf + + output: + file "*.annotatePeaks.txt" into ch_mrep_macs_consensus_annotate + + script: + prefix = "consensus_peaks.mRp.clN" + """ + annotatePeaks.pl \\ + $bed \\ + $fasta \\ + -gid \\ + -gtf $gtf \\ + -cpu $task.cpus \\ + > ${prefix}.annotatePeaks.txt + + cut -f2- ${prefix}.annotatePeaks.txt | awk 'NR==1; NR > 1 {print \$0 | "sort -T '.' -k1,1 -k2,2n"}' | cut -f6- > tmp.txt + paste $bool tmp.txt > ${prefix}.boolean.annotatePeaks.txt + """ +} + +/* + * STEP 8.7 Count reads in consensus peaks with featureCounts + */ +process MergedRepConsensusPeakSetCounts { + label 'process_medium' + publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: 'copy' + + when: + !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks + + input: + file bams from ch_mlib_name_bam_mrep_counts.collect{ it[1] } + file saf from ch_mrep_macs_consensus_saf.collect() + + output: + file "*featureCounts.txt" into ch_mrep_macs_consensus_counts + file "*featureCounts.txt.summary" into ch_mrep_macs_consensus_counts_mqc + + script: + prefix = "consensus_peaks.mRp.clN" + bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() + pe_params = params.single_end ? '' : "-p --donotsort" + """ + featureCounts \\ + -F SAF \\ + -O \\ + --fracOverlap 0.2 \\ + -T $task.cpus \\ + $pe_params \\ + -a $saf \\ + -o ${prefix}.featureCounts.txt \\ + ${bam_files.join(' ')} + """ +} + +/* + * STEP 8.8 Perform differential analysis with DESeq2 + */ +process MergedRepConsensusPeakSetDESeq { + label 'process_medium' + publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus/deseq2", mode: 'copy', + saveAs: { filename -> + if (filename.endsWith(".igv.txt")) null + else filename + } + + when: + !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks && !params.skip_diff_analysis + + input: + file counts from ch_mrep_macs_consensus_counts + file mrep_deseq2_pca_header from ch_mrep_deseq2_pca_header + file mrep_deseq2_clustering_header from ch_mrep_deseq2_clustering_header + + output: + file "*.{RData,results.txt,pdf,log}" into ch_mrep_macs_consensus_deseq_results + file "sizeFactors" into ch_mrep_macs_consensus_deseq_factors + file "*vs*/*.{pdf,txt}" into ch_mrep_macs_consensus_deseq_comp_results + file "*vs*/*.bed" into ch_mrep_macs_consensus_deseq_comp_bed + file "*igv.txt" into ch_mrep_macs_consensus_deseq_comp_igv + file "*.tsv" into ch_mrep_macs_consensus_deseq_mqc + + script: + prefix = "consensus_peaks.mRp.clN" + bam_ext = params.single_end ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" + """ + featurecounts_deseq2.r \\ + --featurecount_file $counts \\ + --bam_suffix '$bam_ext' \\ + --outdir ./ \\ + --outprefix $prefix \\ + --outsuffix .mRp \\ + --cores $task.cpus + + cat $mrep_deseq2_pca_header ${prefix}.pca.vals.txt > ${prefix}.pca.vals_mqc.tsv + cat $mrep_deseq2_clustering_header ${prefix}.sample.dists.txt > ${prefix}.sample.dists_mqc.tsv + + find * -type f -name "*.FDR0.05.results.bed" -exec echo -e "bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus/deseq2/"{}"\\t255,0,0" \\; > ${prefix}.igv.txt + """ +} + +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// +/* -- -- */ +/* -- IGV -- */ +/* -- -- */ +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// + +/* + * STEP 9 - Create IGV session file + */ +process IGV { + publishDir "${params.outdir}/igv/${PEAK_TYPE}", mode: 'copy' + + when: + !params.skip_igv + + input: + file fasta from ch_fasta + + file bigwigs from ch_mlib_bigwig_igv.collect().ifEmpty([]) + file peaks from ch_mlib_macs_igv.collect().ifEmpty([]) + file consensus_peaks from ch_mlib_macs_consensus_igv.collect().ifEmpty([]) + file differential_peaks from ch_mlib_macs_consensus_deseq_comp_igv.collect().ifEmpty([]) + + file rbigwigs from ch_mrep_bigwig_igv.collect().ifEmpty([]) + file rpeaks from ch_mrep_macs_igv.collect().ifEmpty([]) + file rconsensus_peaks from ch_mrep_macs_consensus_igv.collect().ifEmpty([]) + file rdifferential_peaks from ch_mrep_macs_consensus_deseq_comp_igv.collect().ifEmpty([]) + + output: + file "*.{txt,xml}" into ch_igv_session + + script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ + """ + cat *.txt > igv_files.txt + igv_files_to_session.py igv_session.xml igv_files.txt ../../genome/${fasta.getName()} --path_prefix '../../' + """ +} + +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// +/* -- -- */ +/* -- MULTIQC -- */ +/* -- -- */ +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// + +/* + * Parse software version numbers + */ +process get_software_versions { + publishDir "${params.outdir}/pipeline_info", mode: 'copy', + saveAs: { filename -> + if (filename.indexOf(".csv") > 0) filename + else null + } + + output: + file 'software_versions_mqc.yaml' into ch_software_versions_mqc + file "software_versions.csv" + + script: + """ + echo $workflow.manifest.version > v_pipeline.txt + echo $workflow.nextflow.version > v_nextflow.txt + fastqc --version > v_fastqc.txt + trim_galore --version > v_trim_galore.txt + echo \$(bwa 2>&1) > v_bwa.txt + samtools --version > v_samtools.txt + bedtools --version > v_bedtools.txt + echo \$(bamtools --version 2>&1) > v_bamtools.txt + echo \$(plotFingerprint --version 2>&1) > v_deeptools.txt || true + picard MarkDuplicates --version &> v_picard.txt || true + echo \$(R --version 2>&1) > v_R.txt + python -c "import pysam; print(pysam.__version__)" > v_pysam.txt + echo \$(macs2 --version 2>&1) > v_macs2.txt + touch v_homer.txt + echo \$(ataqv --version 2>&1) > v_ataqv.txt + echo \$(featureCounts -v 2>&1) > v_featurecounts.txt + preseq &> v_preseq.txt + multiqc --version > v_multiqc.txt + scrape_software_versions.py &> software_versions_mqc.yaml + """ +} + +Channel.from(summary.collect{ [it.key, it.value] }) + .map { k,v -> "
$k
${v ?: 'N/A'}
" } + .reduce { a, b -> return [a, b].join("\n ") } + .map { x -> """ + id: 'nf-core-chipseq-summary' + description: " - this information is collected when the pipeline is started." + section_name: 'nf-core/chipseq Workflow Summary' + section_href: 'https://github.com/nf-core/chipseq' + plot_type: 'html' + data: | +
+ $x +
+ """.stripIndent() } + .set { ch_workflow_summary } + +/* + * STEP 10 - MultiQC + */ +process MultiQC { + publishDir "${params.outdir}/multiqc/${PEAK_TYPE}", mode: 'copy' + + when: + !params.skip_multiqc + + input: + file (multiqc_config) from ch_multiqc_config + file (mqc_custom_config) from ch_multiqc_custom_config.collect().ifEmpty([]) + + file ('software_versions/*') from ch_software_versions_mqc.collect() + file workflow_summary from ch_workflow_summary.collectFile(name: "workflow_summary_mqc.yaml") + + file ('fastqc/*') from ch_fastqc_reports_mqc.collect().ifEmpty([]) + file ('trimgalore/*') from ch_trimgalore_results_mqc.collect().ifEmpty([]) + file ('trimgalore/fastqc/*') from ch_trimgalore_fastqc_reports_mqc.collect().ifEmpty([]) + + file ('alignment/library/*') from ch_sort_bam_flagstat_mqc.collect() + + file ('alignment/mergedLibrary/*') from ch_mlib_bam_stats_mqc.collect() + file ('alignment/mergedLibrary/*') from ch_mlib_rm_orphan_flagstat_mqc.collect{it[1]} + file ('alignment/mergedLibrary/*') from ch_mlib_rm_orphan_stats_mqc.collect() + file ('alignment/mergedLibrary/picard_metrics/*') from ch_mlib_bam_metrics_mqc.collect() + file ('alignment/mergedLibrary/picard_metrics/*') from ch_mlib_collectmetrics_mqc.collect() + file ('macs/mergedLibrary/*') from ch_mlib_macs_mqc.collect().ifEmpty([]) + file ('macs/mergedLibrary/*') from ch_mlib_peak_qc_mqc.collect().ifEmpty([]) + file ('macs/mergedLibrary/consensus/*') from ch_mlib_macs_consensus_counts_mqc.collect().ifEmpty([]) + file ('macs/mergedLibrary/consensus/*') from ch_mlib_macs_consensus_deseq_mqc.collect().ifEmpty([]) + file ('preseq/*') from ch_mlib_preseq_mqc.collect().ifEmpty([]) + file ('deeptools/*') from ch_mlib_plotprofile_mqc.collect().ifEmpty([]) + file ('deeptools/*') from ch_mlib_plotfingerprint_mqc.collect().ifEmpty([]) + + file ('alignment/mergedReplicate/*') from ch_mrep_bam_flagstat_mqc.collect{it[1]}.ifEmpty([]) + file ('alignment/mergedReplicate/*') from ch_mrep_bam_stats_mqc.collect().ifEmpty([]) + file ('alignment/mergedReplicate/*') from ch_mrep_bam_metrics_mqc.collect().ifEmpty([]) + file ('macs/mergedReplicate/*') from ch_mrep_macs_mqc.collect().ifEmpty([]) + file ('macs/mergedReplicate/*') from ch_mrep_peak_qc_mqc.collect().ifEmpty([]) + file ('macs/mergedReplicate/consensus/*') from ch_mrep_macs_consensus_counts_mqc.collect().ifEmpty([]) + file ('macs/mergedReplicate/consensus/*') from ch_mrep_macs_consensus_deseq_mqc.collect().ifEmpty([]) + + output: + file "*multiqc_report.html" into ch_multiqc_report + file "*_data" + file "multiqc_plots" + + script: + rtitle = custom_runName ? "--title \"$custom_runName\"" : '' + rfilename = custom_runName ? "--filename " + custom_runName.replaceAll('\\W','_').replaceAll('_+','_') + "_multiqc_report" : '' + custom_config_file = params.multiqc_config ? "--config $mqc_custom_config" : '' + """ + multiqc . -f $rtitle $rfilename $custom_config_file \\ + -m custom_content -m fastqc -m cutadapt -m samtools -m picard -m preseq -m featureCounts -m deeptools + """ +} + +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// +/* -- -- */ +/* -- REPORTS/DOCUMENTATION -- */ +/* -- -- */ +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// + +/* + * STEP 11 - Output description HTML + */ +process output_documentation { + publishDir "${params.outdir}/Documentation", mode: 'copy' + + input: + file output_docs from ch_output_docs + + output: + file "results_description.html" + + script: + """ + markdown_to_html.py $output_docs -o results_description.html + """ +} + +/* + * Completion e-mail notification + */ +workflow.onComplete { + + // Set up the e-mail variables + def subject = "[nf-core/atacseq] Successful: $workflow.runName" + if (!workflow.success) { + subject = "[nf-core/atacseq] FAILED: $workflow.runName" + } + def email_fields = [:] + email_fields['version'] = workflow.manifest.version + email_fields['runName'] = custom_runName ?: workflow.runName + email_fields['success'] = workflow.success + email_fields['dateComplete'] = workflow.complete + email_fields['duration'] = workflow.duration + email_fields['exitStatus'] = workflow.exitStatus + email_fields['errorMessage'] = (workflow.errorMessage ?: 'None') + email_fields['errorReport'] = (workflow.errorReport ?: 'None') + email_fields['commandLine'] = workflow.commandLine + email_fields['projectDir'] = workflow.projectDir + email_fields['summary'] = summary + email_fields['summary']['Date Started'] = workflow.start + email_fields['summary']['Date Completed'] = workflow.complete + email_fields['summary']['Pipeline script file path'] = workflow.scriptFile + email_fields['summary']['Pipeline script hash ID'] = workflow.scriptId + if (workflow.repository) email_fields['summary']['Pipeline repository Git URL'] = workflow.repository + if (workflow.commitId) email_fields['summary']['Pipeline repository Git Commit'] = workflow.commitId + if (workflow.revision) email_fields['summary']['Pipeline Git branch/tag'] = workflow.revision + email_fields['summary']['Nextflow Version'] = workflow.nextflow.version + email_fields['summary']['Nextflow Build'] = workflow.nextflow.build + email_fields['summary']['Nextflow Compile Timestamp'] = workflow.nextflow.timestamp + + // On success try attach the multiqc report + def mqc_report = null + try { + if (workflow.success) { + mqc_report = ch_multiqc_report.getVal() + if (mqc_report.getClass() == ArrayList) { + log.warn "[nf-core/atacseq] Found multiple reports from process 'multiqc', will use only one" + mqc_report = mqc_report[0] + } + } + } catch (all) { + log.warn "[nf-core/atacseq] Could not attach MultiQC report to summary email" + } + + // Check if we are only sending emails on failure + email_address = params.email + if (!params.email && params.email_on_fail && !workflow.success) { + email_address = params.email_on_fail + } + + // Render the TXT template + def engine = new groovy.text.GStringTemplateEngine() + def tf = new File("$baseDir/assets/email_template.txt") + def txt_template = engine.createTemplate(tf).make(email_fields) + def email_txt = txt_template.toString() + + // Render the HTML template + def hf = new File("$baseDir/assets/email_template.html") + def html_template = engine.createTemplate(hf).make(email_fields) + def email_html = html_template.toString() + + // Render the sendmail template + def smail_fields = [ email: email_address, subject: subject, email_txt: email_txt, email_html: email_html, baseDir: "$baseDir", mqcFile: mqc_report, mqcMaxSize: params.max_multiqc_email_size.toBytes() ] + def sf = new File("$baseDir/assets/sendmail_template.txt") + def sendmail_template = engine.createTemplate(sf).make(smail_fields) + def sendmail_html = sendmail_template.toString() + + // Send the HTML e-mail + if (email_address) { + try { + if (params.plaintext_email) { throw GroovyException('Send plaintext e-mail, not HTML') } + // Try to send HTML e-mail using sendmail + [ 'sendmail', '-t' ].execute() << sendmail_html + log.info "[nf-core/atacseq] Sent summary e-mail to $email_address (sendmail)" + } catch (all) { + // Catch failures and try with plaintext + [ 'mail', '-s', subject, email_address ].execute() << email_txt + log.info "[nf-core/atacseq] Sent summary e-mail to $email_address (mail)" + } + } + + // Write summary e-mail HTML to a file + def output_d = new File("${params.outdir}/pipeline_info/") + if (!output_d.exists()) { + output_d.mkdirs() + } + def output_hf = new File(output_d, "pipeline_report.html") + output_hf.withWriter { w -> w << email_html } + def output_tf = new File(output_d, "pipeline_report.txt") + output_tf.withWriter { w -> w << email_txt } + + c_green = params.monochrome_logs ? '' : "\033[0;32m"; + c_purple = params.monochrome_logs ? '' : "\033[0;35m"; + c_red = params.monochrome_logs ? '' : "\033[0;31m"; + c_reset = params.monochrome_logs ? '' : "\033[0m"; + + if (workflow.stats.ignoredCount > 0 && workflow.success) { + log.info "-${c_purple}Warning, pipeline completed, but with errored process(es) ${c_reset}-" + log.info "-${c_red}Number of ignored errored process(es) : ${workflow.stats.ignoredCount} ${c_reset}-" + log.info "-${c_green}Number of successfully ran process(es) : ${workflow.stats.succeedCount} ${c_reset}-" + } + if (workflow.success) { + log.info "-${c_purple}[nf-core/atacseq]${c_green} Pipeline completed successfully${c_reset}-" + } else { + checkHostname() + log.info "-${c_purple}[nf-core/atacseq]${c_red} Pipeline completed with errors${c_reset}-" + } + +} /////////////////////////////////////////////////////////////////////////////// /////////////////////////////////////////////////////////////////////////////// From befcb911db5272df8cbb3c483343722f3fece5e0 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Mon, 24 Feb 2020 12:25:59 +0000 Subject: [PATCH 108/226] Update CHANGELOG --- CHANGELOG.md | 4 +++- 1 file changed, 3 insertions(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 126041aa..724036e7 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -8,10 +8,12 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Added` * [#63](https://github.com/nf-core/atacseq/issues/63) - Added multicore support for Trim Galore! +* [#71](https://github.com/nf-core/atacseq/issues/71) - consensus_peaks.mLb.clN.boolean.intersect.plot.pdf not generated * [#75](https://github.com/nf-core/atacseq/issues/75) - Include gene annotation versions in multiqc report * [#76](https://github.com/nf-core/atacseq/issues/76) - featureCounts coupled to DESeq2 -* [79](https://github.com/nf-core/atacseq/issues/79) - Parallelize DESeq2 +* [#79](https://github.com/nf-core/atacseq/issues/79) - Parallelize DESeq2 * [#80](https://github.com/nf-core/atacseq/pull/80) - Added social preview image + * Update template to tools `1.9` * Parameter `--skip_consensus_peaks` to skip consensus peak generation From 36b212fa51fade3aecda57192967f73ec7a55816 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Mon, 24 Feb 2020 12:26:12 +0000 Subject: [PATCH 109/226] Change ataqv suffix --- main.nf | 7 ++++--- 1 file changed, 4 insertions(+), 3 deletions(-) diff --git a/main.nf b/main.nf index 2df57e44..dbe5900f 100644 --- a/main.nf +++ b/main.nf @@ -1351,7 +1351,7 @@ process MergedLibConsensusPeakSetDESeq { --bam_suffix '$bam_ext' \\ --outdir ./ \\ --outprefix $prefix \\ - --outsuffix .mLb \\ + --outsuffix .mLb.clN \\ --cores $task.cpus cat $mlib_deseq2_pca_header ${prefix}.pca.vals.txt > ${prefix}.pca.vals_mqc.tsv @@ -1381,6 +1381,7 @@ process MergedLibAtaqv { file "*.json" into ch_mlib_ataqv script: + suffix = 'mLb.clN' peak_param = params.macs_gsize ? "--peak-file ${peak}" : "" mito_param = params.mito_name ? "--mitochondrial-reference-name ${params.mito_name}" : "" """ @@ -1388,7 +1389,7 @@ process MergedLibAtaqv { --threads $task.cpus \\ $peak_param \\ --tss-file $tss_bed \\ - --metrics-file ${name}.ataqv.json \\ + --metrics-file ${name}.${suffix}.ataqv.json \\ --name $name \\ --ignore-read-groups \\ --autosomal-reference-file $autosomes \\ @@ -1843,7 +1844,7 @@ process MergedRepConsensusPeakSetDESeq { --bam_suffix '$bam_ext' \\ --outdir ./ \\ --outprefix $prefix \\ - --outsuffix .mRp \\ + --outsuffix .mRp.clN \\ --cores $task.cpus cat $mrep_deseq2_pca_header ${prefix}.pca.vals.txt > ${prefix}.pca.vals_mqc.tsv From d6546e38abd5c6f01be129be78bba43e835f9e5c Mon Sep 17 00:00:00 2001 From: drpatelh Date: Mon, 24 Feb 2020 12:43:40 +0000 Subject: [PATCH 110/226] Argggh copy paste --- main.nf | 6 +++--- 1 file changed, 3 insertions(+), 3 deletions(-) diff --git a/main.nf b/main.nf index dbe5900f..27e6dfa3 100644 --- a/main.nf +++ b/main.nf @@ -1944,10 +1944,10 @@ Channel.from(summary.collect{ [it.key, it.value] }) .map { k,v -> "
$k
${v ?: 'N/A'}
" } .reduce { a, b -> return [a, b].join("\n ") } .map { x -> """ - id: 'nf-core-chipseq-summary' + id: 'nf-core-atacseq-summary' description: " - this information is collected when the pipeline is started." - section_name: 'nf-core/chipseq Workflow Summary' - section_href: 'https://github.com/nf-core/chipseq' + section_name: 'nf-core/atacseq Workflow Summary' + section_href: 'https://github.com/nf-core/atacseq' plot_type: 'html' data: |
From 6aed3ccf2d2e8233ced5adc26ef406ddb7864e41 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Mon, 24 Feb 2020 14:26:27 +0000 Subject: [PATCH 111/226] Consistent metavar definitions --- bin/plot_homer_annotatepeaks.r | 6 +++--- bin/plot_macs_qc.r | 6 +++--- bin/plot_peak_intersect.r | 4 ++-- 3 files changed, 8 insertions(+), 8 deletions(-) diff --git a/bin/plot_homer_annotatepeaks.r b/bin/plot_homer_annotatepeaks.r index 57ca4b67..6219ae28 100755 --- a/bin/plot_homer_annotatepeaks.r +++ b/bin/plot_homer_annotatepeaks.r @@ -17,10 +17,10 @@ library(scales) ################################################ ################################################ -option_list <- list(make_option(c("-i", "--homer_files"), type="character", default=NULL, help="Comma-separated list of homer annotated text files.", metavar="anno_files"), - make_option(c("-s", "--sample_ids"), type="character", default=NULL, help="Comma-separated list of sample ids associated with homer annotated text files. Must be unique and in same order as homer files input.", metavar="sampleids"), +option_list <- list(make_option(c("-i", "--homer_files"), type="character", default=NULL, help="Comma-separated list of homer annotated text files.", metavar="path"), + make_option(c("-s", "--sample_ids"), type="character", default=NULL, help="Comma-separated list of sample ids associated with homer annotated text files. Must be unique and in same order as homer files input.", metavar="string"), make_option(c("-o", "--outdir"), type="character", default='./', help="Output directory", metavar="path"), - make_option(c("-p", "--outprefix"), type="character", default='homer_annotation', help="Output prefix", metavar="character")) + make_option(c("-p", "--outprefix"), type="character", default='homer_annotation', help="Output prefix", metavar="string")) opt_parser <- OptionParser(option_list=option_list) opt <- parse_args(opt_parser) diff --git a/bin/plot_macs_qc.r b/bin/plot_macs_qc.r index 2360505f..b8e25d56 100755 --- a/bin/plot_macs_qc.r +++ b/bin/plot_macs_qc.r @@ -17,10 +17,10 @@ library(scales) ################################################ ################################################ -option_list <- list(make_option(c("-i", "--peak_files"), type="character", default=NULL, help="Comma-separated list of peak files.", metavar="peak_files"), - make_option(c("-s", "--sample_ids"), type="character", default=NULL, help="Comma-separated list of sample ids associated with peak files. Must be unique and in same order as peaks files input.", metavar="sampleids"), +option_list <- list(make_option(c("-i", "--peak_files"), type="character", default=NULL, help="Comma-separated list of peak files.", metavar="path"), + make_option(c("-s", "--sample_ids"), type="character", default=NULL, help="Comma-separated list of sample ids associated with peak files. Must be unique and in same order as peaks files input.", metavar="string"), make_option(c("-o", "--outdir"), type="character", default='./', help="Output directory", metavar="path"), - make_option(c("-p", "--outprefix"), type="character", default='macs2_peakqc', help="Output prefix", metavar="character")) + make_option(c("-p", "--outprefix"), type="character", default='macs2_peakqc', help="Output prefix", metavar="string")) opt_parser <- OptionParser(option_list=option_list) opt <- parse_args(opt_parser) diff --git a/bin/plot_peak_intersect.r b/bin/plot_peak_intersect.r index 40abf6d3..513e44b3 100755 --- a/bin/plot_peak_intersect.r +++ b/bin/plot_peak_intersect.r @@ -15,8 +15,8 @@ library(UpSetR) ################################################ ################################################ -option_list <- list(make_option(c("-i", "--input_file"), type="character", default=NULL, help="Path to tab-delimited file containing two columns i.e sample1&sample2&sample3 indicating intersect between samples set size.", metavar="input_file"), - make_option(c("-o", "--output_file"), type="character", default=NULL, help="Path to output file with '.pdf' extension.", metavar="output_file")) +option_list <- list(make_option(c("-i", "--input_file"), type="character", default=NULL, help="Path to tab-delimited file containing two columns i.e sample1&sample2&sample3 indicating intersect between samples set size.", metavar="path"), + make_option(c("-o", "--output_file"), type="character", default=NULL, help="Path to output file with '.pdf' extension.", metavar="path")) opt_parser <- OptionParser(option_list=option_list) opt <- parse_args(opt_parser) From 5dc2eb974082a74193f9a617805c28d0e29671ba Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 11 Mar 2020 11:15:47 +0000 Subject: [PATCH 112/226] Create empty .Rprofile in Dockerfile --- Dockerfile | 3 +++ 1 file changed, 3 insertions(+) diff --git a/Dockerfile b/Dockerfile index e0478eae..55dc5359 100644 --- a/Dockerfile +++ b/Dockerfile @@ -11,3 +11,6 @@ ENV PATH /opt/conda/envs/nf-core-atacseq-1.1.1dev/bin:$PATH # Dump the details of the installed packages to a file for posterity RUN conda env export --name nf-core-atacseq-1.1.1dev > nf-core-atacseq-1.1.1dev.yml + +# Instruct R processes to use this empty file instead of clashing with a local version +RUN touch .Rprofile From 3efa0f111a5b66c9a58c44d17ebffe1159938c79 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 11 Mar 2020 11:17:56 +0000 Subject: [PATCH 113/226] Export R_PROFILE_USER in env scope --- nextflow.config | 3 ++- 1 file changed, 2 insertions(+), 1 deletion(-) diff --git a/nextflow.config b/nextflow.config index e5400b59..5d4b32c5 100755 --- a/nextflow.config +++ b/nextflow.config @@ -127,9 +127,10 @@ if (!params.igenomes_ignore) { // Increase time available to build conda environment conda { createTimeout = "60 min" } -// Export this variable to prevent local Python libraries from conflicting with those in the container +// Export these variables to prevent local Python/R libraries from conflicting with those in the container env { PYTHONNOUSERSITE = 1 + R_PROFILE_USER=~/.Rprofile } // Capture exit codes from upstream processes when piping From 9d780ee23d0923404b759ccf2de03021e54cc9ce Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 11 Mar 2020 11:21:08 +0000 Subject: [PATCH 114/226] Add spaces --- nextflow.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/nextflow.config b/nextflow.config index 5d4b32c5..9df96d6b 100755 --- a/nextflow.config +++ b/nextflow.config @@ -130,7 +130,7 @@ conda { createTimeout = "60 min" } // Export these variables to prevent local Python/R libraries from conflicting with those in the container env { PYTHONNOUSERSITE = 1 - R_PROFILE_USER=~/.Rprofile + R_PROFILE_USER = ~/.Rprofile } // Capture exit codes from upstream processes when piping From e55663678854a97de6dc1125ad79f9f82da54ac9 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 11 Mar 2020 11:36:37 +0000 Subject: [PATCH 115/226] Add quotes --- nextflow.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/nextflow.config b/nextflow.config index 9df96d6b..32b9e1ff 100755 --- a/nextflow.config +++ b/nextflow.config @@ -130,7 +130,7 @@ conda { createTimeout = "60 min" } // Export these variables to prevent local Python/R libraries from conflicting with those in the container env { PYTHONNOUSERSITE = 1 - R_PROFILE_USER = ~/.Rprofile + R_PROFILE_USER = "~/.Rprofile" } // Capture exit codes from upstream processes when piping From cf6c5c39d890a3a69afa42b9316db02670b85f8b Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 11 Mar 2020 11:43:18 +0000 Subject: [PATCH 116/226] Replace read.table with read.csv --- bin/plot_homer_annotatepeaks.r | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/bin/plot_homer_annotatepeaks.r b/bin/plot_homer_annotatepeaks.r index 6219ae28..4a867d8f 100755 --- a/bin/plot_homer_annotatepeaks.r +++ b/bin/plot_homer_annotatepeaks.r @@ -57,7 +57,7 @@ plot.feature.dat <- data.frame() for (idx in 1:length(HomerFiles)) { sampleid = SampleIDs[idx] - anno.dat <- read.table(HomerFiles[idx], sep="\t", header=TRUE,quote="") + anno.dat <- read.csv(HomerFiles[idx], sep="\t", header=TRUE) anno.dat <- anno.dat[,c("Annotation","Distance.to.TSS","Nearest.PromoterID")] ## REPLACE UNASSIGNED FEATURE ENTRIES WITH SENSIBLE VALUES From 162926716ffee5a466ea8db2e41eee98ae1aea1f Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 11 Mar 2020 11:50:26 +0000 Subject: [PATCH 117/226] Update CHANGELOG --- CHANGELOG.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 724036e7..494a8b20 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -13,7 +13,6 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * [#76](https://github.com/nf-core/atacseq/issues/76) - featureCounts coupled to DESeq2 * [#79](https://github.com/nf-core/atacseq/issues/79) - Parallelize DESeq2 * [#80](https://github.com/nf-core/atacseq/pull/80) - Added social preview image - * Update template to tools `1.9` * Parameter `--skip_consensus_peaks` to skip consensus peak generation @@ -22,6 +21,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * [nf-core/chipseq#118](https://github.com/nf-core/chipseq/issues/118) - Running on with SGE * [nf-core/chipseq#132](https://github.com/nf-core/chipseq/issues/132) - BigWig Error: sort: cannot create temporary file in '': Read-only file system * [#73](https://github.com/nf-core/atacseq/issues/73) - macs_annotatePeaks.mLb.clN.summary.txt file is not created +* [#86](https://github.com/nf-core/atacseq/issues/86) - bug in the plot_homer_annotatepeaks.r script * Make executables in `bin/` compatible with Python 3 ### `Dependencies` From 2defdec83e1008d6f88a2e6bd7c6072239bdea91 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 11 Mar 2020 15:39:34 +0000 Subject: [PATCH 118/226] Update CHANGELOG --- CHANGELOG.md | 4 +++- 1 file changed, 3 insertions(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 494a8b20..1c775b1d 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -14,7 +14,9 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * [#79](https://github.com/nf-core/atacseq/issues/79) - Parallelize DESeq2 * [#80](https://github.com/nf-core/atacseq/pull/80) - Added social preview image * Update template to tools `1.9` -* Parameter `--skip_consensus_peaks` to skip consensus peak generation +* Parameters: + * `--skip_consensus_peaks` to skip consensus peak generation + * `--deseq2_vst` to use `vst` transformation instead of `rlog` with DESeq2 ### `Fixed` From 806f772363f7a330eee031e80b5381a17b23c011 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 11 Mar 2020 15:39:42 +0000 Subject: [PATCH 119/226] Add .Renviron --- Dockerfile | 3 ++- 1 file changed, 2 insertions(+), 1 deletion(-) diff --git a/Dockerfile b/Dockerfile index 55dc5359..ecae8934 100644 --- a/Dockerfile +++ b/Dockerfile @@ -12,5 +12,6 @@ ENV PATH /opt/conda/envs/nf-core-atacseq-1.1.1dev/bin:$PATH # Dump the details of the installed packages to a file for posterity RUN conda env export --name nf-core-atacseq-1.1.1dev > nf-core-atacseq-1.1.1dev.yml -# Instruct R processes to use this empty file instead of clashing with a local version +# Instruct R processes to use these empty files instead of clashing with a local version RUN touch .Rprofile +RUN touch .Renviron From 1a31d9d07cd62deb98a5bd1694f4a3ff3e5246f8 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 11 Mar 2020 15:39:46 +0000 Subject: [PATCH 120/226] Add .Renviron --- nextflow.config | 4 ++++ 1 file changed, 4 insertions(+) diff --git a/nextflow.config b/nextflow.config index 32b9e1ff..f2485777 100755 --- a/nextflow.config +++ b/nextflow.config @@ -41,6 +41,9 @@ params { min_reps_consensus = 1 save_macs_pileup = false skip_consensus_peaks = false + + // Options: Differential analysis + deseq2_vst = false skip_diff_analysis = false // Options: QC @@ -131,6 +134,7 @@ conda { createTimeout = "60 min" } env { PYTHONNOUSERSITE = 1 R_PROFILE_USER = "~/.Rprofile" + R_ENVIRON_USER = "~/.Renviron" } // Capture exit codes from upstream processes when piping From eff61ea2f91e2968fa8502aa81e9318d6e8fdbe4 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 11 Mar 2020 15:39:59 +0000 Subject: [PATCH 121/226] Add vsn parameter --- bin/featurecounts_deseq2.r | 7 ++++++- 1 file changed, 6 insertions(+), 1 deletion(-) diff --git a/bin/featurecounts_deseq2.r b/bin/featurecounts_deseq2.r index d9c07a0f..e19fd977 100755 --- a/bin/featurecounts_deseq2.r +++ b/bin/featurecounts_deseq2.r @@ -38,6 +38,7 @@ option_list <- list(make_option(c("-i", "--featurecount_file"), type="character" make_option(c("-o", "--outdir"), type="character", default='./', help="Output directory", metavar="path"), make_option(c("-p", "--outprefix"), type="character", default='differential', help="Output prefix", metavar="string"), make_option(c("-s", "--outsuffix"), type="character", default='', help="Output suffix for comparison-level results", metavar="string"), + make_option(c("-v", "--vst"), type="logical", default=FALSE, help="Run vst transform instead of rlog", metavar="boolean"), make_option(c("-c", "--cores"), type="integer", default=1, help="Number of cores", metavar="integer")) opt_parser <- OptionParser(option_list=option_list) @@ -89,7 +90,11 @@ if (file.exists(DDSFile) == FALSE) { coldata <- data.frame(row.names=colnames(counts),condition=groups) dds <- DESeqDataSetFromMatrix(countData = round(counts), colData = coldata, design = ~ condition) dds <- DESeq(dds, parallel=TRUE, BPPARAM=MulticoreParam(opt$cores)) - rld <- rlog(dds) + if (!opt$vst) { + rld <- rlog(dds) + } else { + rld <- vst(dds) + } save(dds,rld,file=DDSFile) } From 657e559042cb282cc4f821f3ddc5c793574bcd8d Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 11 Mar 2020 15:40:13 +0000 Subject: [PATCH 122/226] Add deseq2_vsn parameter --- docs/output.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/docs/output.md b/docs/output.md index 59fbf36b..c35eda84 100755 --- a/docs/output.md +++ b/docs/output.md @@ -210,7 +210,7 @@ The library-level alignments associated with the same sample are merged and subs *Description*: DESeq2 is more commonly used to perform differential expression analysis for RNA-seq datasets. However, it can also be used for ATAC-seq differential accessibility analysis, in which case you can imagine that instead of counts per gene for RNA-seq data we now have counts per accessible region. - This pipeline uses a standardised DESeq2 analysis script to get an idea of the reproducibility within the experiment, and to assess the overall differential binding. Please note that this will not suit every experimental design, and if there are other problems with the experiment then it may not work as well as expected. + This pipeline uses a standardised DESeq2 analysis script to get an idea of the reproducibility within the experiment, and to assess the overall differential binding. Please note that this will not suit every experimental design, and if there are other problems with the experiment then it may not work as well as expected. For larger experiments, it may be recommended to use the `vst` transformation instead of the default `rlog` option. You can do this by providing the `--deseq2_vsn` parameter to the pipeline. ![MultiQC - DESeq2 PCA plot](images/mqc_deseq2_pca_plot.png) From e3b7543a059bce8b7b0253954f688cdb5adae737 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 11 Mar 2020 15:40:19 +0000 Subject: [PATCH 123/226] Add deseq2_vsn parameter --- docs/usage.md | 8 ++++++++ 1 file changed, 8 insertions(+) diff --git a/docs/usage.md b/docs/usage.md index 9e689f68..21ed1171 100755 --- a/docs/usage.md +++ b/docs/usage.md @@ -42,6 +42,8 @@ * [`--min_reps_consensus`](#--min_reps_consensus) * [`--save_macs_pileup`](#--save_macs_pileup) * [`--skip_consensus_peaks`](#--skip_consensus_peaks) +* [Differential analysis](#differential_analysis) + * [`--deseq2_vst`](#--deseq2_vst) * [`--skip_diff_analysis`](#--skip_diff_analysis) * [Skipping QC steps](#skipping-qc-steps) * [Job resources](#job-resources) @@ -399,6 +401,12 @@ Instruct MACS2 to create bedGraph files using the `-B --SPMR` parameters. Skip consensus peak generation, annotation and counting. +## Differential analysis + +### `--deseq2_vst` + +Use `vst` transformation instead of `rlog` with DESeq2. See [DESeq2 docs](http://bioconductor.org/packages/devel/bioc/vignettes/DESeq2/inst/doc/DESeq2.html#data-transformations-and-visualization). + ### `--skip_diff_analysis` Skip differential accessibility analysis with DESeq2. From ccdd4a73492407dd097c2e739e47952efa80569e Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 11 Mar 2020 15:40:33 +0000 Subject: [PATCH 124/226] Add deseq2_vsn parameter --- main.nf | 12 ++++++++++-- 1 file changed, 10 insertions(+), 2 deletions(-) diff --git a/main.nf b/main.nf index 27e6dfa3..0bcbcee2 100644 --- a/main.nf +++ b/main.nf @@ -63,6 +63,9 @@ def helpMessage() { --min_reps_consensus [int] Number of biological replicates required from a given condition for a peak to contribute to a consensus peak (Default: 1) --save_macs_pileup [bool] Instruct MACS2 to create bedGraph files normalised to signal per million reads --skip_consensus_peaks [bool] Skip consensus peak generation + + Differential analysis + --deseq2_vst [bool] Use vst transformation instead of rlog with DESeq2 --skip_diff_analysis [bool] Skip differential accessibility analysis QC @@ -260,6 +263,7 @@ if (params.save_align_intermeds) summary['Save Intermeds'] = 'Yes' if (params.save_macs_pileup) summary['Save MACS2 Pileup'] = 'Yes' if (params.skip_merge_replicates) summary['Skip Merge Replicates'] = 'Yes' if (params.skip_consensus_peaks) summary['Skip Consensus Peaks'] = 'Yes' +if (params.deseq2_vst) summary['Use DESeq2 vst Transform'] = 'Yes' if (params.skip_diff_analysis) summary['Skip Differential Analysis'] = 'Yes' if (params.skip_fastqc) summary['Skip FastQC'] = 'Yes' if (params.skip_picard_metrics) summary['Skip Picard Metrics'] = 'Yes' @@ -1345,6 +1349,7 @@ process MergedLibConsensusPeakSetDESeq { script: prefix = "consensus_peaks.mLb.clN" bam_ext = params.single_end ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" + vst = params.deseq2_vst ? "--vst TRUE" : "" """ featurecounts_deseq2.r \\ --featurecount_file $counts \\ @@ -1352,7 +1357,8 @@ process MergedLibConsensusPeakSetDESeq { --outdir ./ \\ --outprefix $prefix \\ --outsuffix .mLb.clN \\ - --cores $task.cpus + --cores $task.cpus \\ + $vst \\ cat $mlib_deseq2_pca_header ${prefix}.pca.vals.txt > ${prefix}.pca.vals_mqc.tsv cat $mlib_deseq2_clustering_header ${prefix}.sample.dists.txt > ${prefix}.sample.dists_mqc.tsv @@ -1838,6 +1844,7 @@ process MergedRepConsensusPeakSetDESeq { script: prefix = "consensus_peaks.mRp.clN" bam_ext = params.single_end ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" + vst = params.deseq2_vst ? "--vst TRUE" : "" """ featurecounts_deseq2.r \\ --featurecount_file $counts \\ @@ -1845,7 +1852,8 @@ process MergedRepConsensusPeakSetDESeq { --outdir ./ \\ --outprefix $prefix \\ --outsuffix .mRp.clN \\ - --cores $task.cpus + --cores $task.cpus \\ + $vst cat $mrep_deseq2_pca_header ${prefix}.pca.vals.txt > ${prefix}.pca.vals_mqc.tsv cat $mrep_deseq2_clustering_header ${prefix}.sample.dists.txt > ${prefix}.sample.dists_mqc.tsv From e8f1584185547abaae1de14f2eb044de94753a48 Mon Sep 17 00:00:00 2001 From: mashehu Date: Wed, 11 Mar 2020 17:13:22 +0100 Subject: [PATCH 125/226] fix typo --- docs/output.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/docs/output.md b/docs/output.md index c35eda84..57df45a0 100755 --- a/docs/output.md +++ b/docs/output.md @@ -210,7 +210,7 @@ The library-level alignments associated with the same sample are merged and subs *Description*: DESeq2 is more commonly used to perform differential expression analysis for RNA-seq datasets. However, it can also be used for ATAC-seq differential accessibility analysis, in which case you can imagine that instead of counts per gene for RNA-seq data we now have counts per accessible region. - This pipeline uses a standardised DESeq2 analysis script to get an idea of the reproducibility within the experiment, and to assess the overall differential binding. Please note that this will not suit every experimental design, and if there are other problems with the experiment then it may not work as well as expected. For larger experiments, it may be recommended to use the `vst` transformation instead of the default `rlog` option. You can do this by providing the `--deseq2_vsn` parameter to the pipeline. + This pipeline uses a standardised DESeq2 analysis script to get an idea of the reproducibility within the experiment, and to assess the overall differential binding. Please note that this will not suit every experimental design, and if there are other problems with the experiment then it may not work as well as expected. For larger experiments, it may be recommended to use the `vst` transformation instead of the default `rlog` option. You can do this by providing the `--deseq2_vst` parameter to the pipeline. ![MultiQC - DESeq2 PCA plot](images/mqc_deseq2_pca_plot.png) From a1516de9689401afd84e5a2617d08ba539e9765f Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 11 Mar 2020 18:25:09 +0000 Subject: [PATCH 126/226] Update CHANGELOG --- CHANGELOG.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 1c775b1d..309e36c7 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -16,7 +16,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * Update template to tools `1.9` * Parameters: * `--skip_consensus_peaks` to skip consensus peak generation - * `--deseq2_vst` to use `vst` transformation instead of `rlog` with DESeq2 + * `--deseq2_vst` to use variance stabilizing transformation (VST) instead of regularized log transformation (rlog) with DESeq2 ### `Fixed` From 62177aa596ad80a1c4d39a0c05968f6ca917b9f6 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 11 Mar 2020 18:43:08 +0000 Subject: [PATCH 127/226] Update contributors --- README.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/README.md b/README.md index 24153332..aa81ccc8 100644 --- a/README.md +++ b/README.md @@ -100,7 +100,7 @@ The pipeline was developed by [Harshil Patel](mailto:harshil.patel@crick.ac.uk). The [nf-core/rnaseq](https://github.com/nf-core/rnaseq) and [nf-core/chipseq](https://github.com/nf-core/chipseq) pipelines developed by Phil Ewels were initially used as a template for this pipeline. Many thanks to Phil for all of his help and advice, and the team at SciLifeLab. -Many thanks to others who have helped out along the way too, including (but not limited to): [@apeltzer](https://github.com/apeltzer), [@sven1103](https://github.com/sven1103), [@MaxUlysse](https://github.com/MaxUlysse), [@micans](https://github.com/micans), [@jinmingda](https://github.com/jinmingda), [@ktrns](https://github.com/ktrns), [@crickbabs](https://github.com/crickbabs), [@pditommaso](https://github.com/pditommaso). +Many thanks to others who have helped out and contributed along the way too, including (but not limited to): [@apeltzer](https://github.com/apeltzer), [@crickbabs](https://github.com/crickbabs), [drewjbeh](https://github.com/drewjbeh), [@houghtos](https://github.com/houghtos), [@jinmingda](https://github.com/jinmingda), [@ktrns](https://github.com/ktrns), [@MaxUlysse](https://github.com/MaxUlysse), [@mashehu](https://github.com/mashehu), [@micans](https://github.com/micans), [@pditommaso](https://github.com/pditommaso) and [@sven1103](https://github.com/sven1103). ## Contributions and Support From 391ae6ad9b277d8f3b26f3c944e2059bcac0c7a5 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 11 Mar 2020 18:43:19 +0000 Subject: [PATCH 128/226] Link to vst docs --- docs/output.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/docs/output.md b/docs/output.md index 57df45a0..b1a4a71c 100755 --- a/docs/output.md +++ b/docs/output.md @@ -210,7 +210,7 @@ The library-level alignments associated with the same sample are merged and subs *Description*: DESeq2 is more commonly used to perform differential expression analysis for RNA-seq datasets. However, it can also be used for ATAC-seq differential accessibility analysis, in which case you can imagine that instead of counts per gene for RNA-seq data we now have counts per accessible region. - This pipeline uses a standardised DESeq2 analysis script to get an idea of the reproducibility within the experiment, and to assess the overall differential binding. Please note that this will not suit every experimental design, and if there are other problems with the experiment then it may not work as well as expected. For larger experiments, it may be recommended to use the `vst` transformation instead of the default `rlog` option. You can do this by providing the `--deseq2_vst` parameter to the pipeline. + This pipeline uses a standardised DESeq2 analysis script to get an idea of the reproducibility within the experiment, and to assess the overall differential binding. Please note that this will not suit every experimental design, and if there are other problems with the experiment then it may not work as well as expected. For larger experiments, it may be recommended to use the `vst` transformation instead of the default `rlog` option. You can do this by providing the `--deseq2_vst` parameter to the pipeline. See [DESeq2 docs](http://bioconductor.org/packages/devel/bioc/vignettes/DESeq2/inst/doc/DESeq2.html#data-transformations-and-visualization) for a more detailed explanation. ![MultiQC - DESeq2 PCA plot](images/mqc_deseq2_pca_plot.png) From 73737141939b8aa0f4c7f27a780fd2d1d9a56530 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 12 Mar 2020 11:33:41 +0000 Subject: [PATCH 129/226] Update Docker citation --- CITATIONS.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/CITATIONS.md b/CITATIONS.md index 68e539f1..2ef37fed 100644 --- a/CITATIONS.md +++ b/CITATIONS.md @@ -97,4 +97,4 @@ * [Singularity](https://www.ncbi.nlm.nih.gov/pubmed/28494014/) > Kurtzer GM, Sochat V, Bauer MW. Singularity: Scientific containers for mobility of compute. PLoS One. 2017 May 11;12(5):e0177459. doi: 10.1371/journal.pone.0177459. eCollection 2017. PubMed PMID: 28494014; PubMed Central PMCID: PMC5426675. -* [Docker](https://www.docker.com/) +* [Docker](https://dl.acm.org/doi/10.5555/2600239.2600241) From 7ccb24ac5da31b02fb5fe2ead7197f862624f0c3 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 12 Mar 2020 13:59:30 +0000 Subject: [PATCH 130/226] Change home to root --- nextflow.config | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/nextflow.config b/nextflow.config index f2485777..6c8fe99c 100755 --- a/nextflow.config +++ b/nextflow.config @@ -133,8 +133,8 @@ conda { createTimeout = "60 min" } // Export these variables to prevent local Python/R libraries from conflicting with those in the container env { PYTHONNOUSERSITE = 1 - R_PROFILE_USER = "~/.Rprofile" - R_ENVIRON_USER = "~/.Renviron" + R_PROFILE_USER = "/.Rprofile" + R_ENVIRON_USER = "/.Renviron" } // Capture exit codes from upstream processes when piping From f86556c8e1d305aee4141833985be6a1c0517113 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 13 Mar 2020 14:41:25 +0000 Subject: [PATCH 131/226] Bump version to 1.2.0 --- .github/workflows/ci.yml | 2 +- Dockerfile | 4 ++-- environment.yml | 2 +- nextflow.config | 4 ++-- 4 files changed, 6 insertions(+), 6 deletions(-) diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml index a442aa32..2f69d7c5 100644 --- a/.github/workflows/ci.yml +++ b/.github/workflows/ci.yml @@ -22,7 +22,7 @@ jobs: - name: Pull docker image run: | docker pull nfcore/atacseq:dev - docker tag nfcore/atacseq:dev nfcore/atacseq:dev + docker tag nfcore/atacseq:dev nfcore/atacseq:1.2.0 - name: Run pipeline with test data run: | nextflow run ${GITHUB_WORKSPACE} -profile test,docker diff --git a/Dockerfile b/Dockerfile index ecae8934..715e6892 100644 --- a/Dockerfile +++ b/Dockerfile @@ -7,10 +7,10 @@ COPY environment.yml / RUN conda env create -f /environment.yml && conda clean -a # Add conda installation dir to PATH (instead of doing 'conda activate') -ENV PATH /opt/conda/envs/nf-core-atacseq-1.1.1dev/bin:$PATH +ENV PATH /opt/conda/envs/nf-core-atacseq-1.2.0/bin:$PATH # Dump the details of the installed packages to a file for posterity -RUN conda env export --name nf-core-atacseq-1.1.1dev > nf-core-atacseq-1.1.1dev.yml +RUN conda env export --name nf-core-atacseq-1.2.0 > nf-core-atacseq-1.2.0.yml # Instruct R processes to use these empty files instead of clashing with a local version RUN touch .Rprofile diff --git a/environment.yml b/environment.yml index af38f938..535b7768 100755 --- a/environment.yml +++ b/environment.yml @@ -1,6 +1,6 @@ # You can use this file to create a conda environment for this pipeline: # conda env create -f environment.yml -name: nf-core-atacseq-1.1.1dev +name: nf-core-atacseq-1.2.0 channels: - conda-forge - bioconda diff --git a/nextflow.config b/nextflow.config index 6c8fe99c..97d2c64c 100755 --- a/nextflow.config +++ b/nextflow.config @@ -92,7 +92,7 @@ params { // Container slug. Stable releases should specify release tag! // Developmental code should specify :dev -process.container = 'nfcore/atacseq:dev' +process.container = 'nfcore/atacseq:1.2.0' // Load base.config by default for all pipelines includeConfig 'conf/base.config' @@ -164,7 +164,7 @@ manifest { description = 'ATACSeq peak-calling and differential analysis pipeline.' mainScript = 'main.nf' nextflowVersion = '>=19.10.0' - version = '1.1.1dev' + version = '1.2.0' } // Function to ensure that resource requirements don't go beyond From af4502f9d749cfca34bfb2dfea0920dbadfe55b3 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 13 Mar 2020 21:50:50 +0000 Subject: [PATCH 132/226] Change data in CHANGELOG --- CHANGELOG.md | 4 +--- 1 file changed, 1 insertion(+), 3 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 309e36c7..5d65c7eb 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -3,7 +3,7 @@ The format is based on [Keep a Changelog](http://keepachangelog.com/en/1.0.0/) and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0.html). -## [Unpublished Version / DEV] +## [1.2.0] - 2020-03-20 ### `Added` @@ -60,8 +60,6 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * Update bioconductor-vsn `3.46.0` -> `3.54.0` * Remove r-reshape2 `1.4.3` -### `Deprecated` - ## [1.1.0] - 2019-11-05 ### `Added` From 0f52c525f1e3d414988a1eea36e28390c348a004 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sun, 15 Mar 2020 19:02:36 +0000 Subject: [PATCH 133/226] Replace with bedgraphToBigWig --- README.md | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/README.md b/README.md index aa81ccc8..d9ae7a95 100644 --- a/README.md +++ b/README.md @@ -36,7 +36,7 @@ The pipeline is built using [Nextflow](https://www.nextflow.io), a workflow tool * reads that arent in FR orientation ([`Pysam`](http://pysam.readthedocs.io/en/latest/installation.html); *paired-end only*) * reads where only one read of the pair fails the above criteria ([`Pysam`](http://pysam.readthedocs.io/en/latest/installation.html); *paired-end only*) 3. Alignment-level QC and estimation of library complexity ([`picard`](https://broadinstitute.github.io/picard/), [`Preseq`](http://smithlabresearch.org/software/preseq/)) - 4. Create normalised bigWig files scaled to 1 million mapped reads ([`BEDTools`](https://github.com/arq5x/bedtools2/), [`wigToBigWig`](http://hgdownload.soe.ucsc.edu/admin/exe/)) + 4. Create normalised bigWig files scaled to 1 million mapped reads ([`BEDTools`](https://github.com/arq5x/bedtools2/), [`bedGraphToBigWig`](http://hgdownload.soe.ucsc.edu/admin/exe/)) 5. Generate gene-body meta-profile from bigWig files ([`deepTools`](https://deeptools.readthedocs.io/en/develop/content/tools/plotProfile.html)) 6. Calculate genome-wide enrichment ([`deepTools`](https://deeptools.readthedocs.io/en/develop/content/tools/plotFingerprint.html)) 7. Call broad/narrow peaks ([`MACS2`](https://github.com/taoliu/MACS)) @@ -48,7 +48,7 @@ The pipeline is built using [Nextflow](https://www.nextflow.io), a workflow tool 6. Merge filtered alignments across replicates ([`picard`](https://broadinstitute.github.io/picard/)) 1. Re-mark duplicates ([`picard`](https://broadinstitute.github.io/picard/)) 2. Remove duplicate reads ([`SAMtools`](https://sourceforge.net/projects/samtools/files/samtools/)) - 3. Create normalised bigWig files scaled to 1 million mapped reads ([`BEDTools`](https://github.com/arq5x/bedtools2/), [`wigToBigWig`](http://hgdownload.soe.ucsc.edu/admin/exe/)) + 3. Create normalised bigWig files scaled to 1 million mapped reads ([`BEDTools`](https://github.com/arq5x/bedtools2/), [`bedGraphToBigWig`](http://hgdownload.soe.ucsc.edu/admin/exe/)) 4. Call broad/narrow peaks ([`MACS2`](https://github.com/taoliu/MACS)) 5. Annotate peaks relative to gene features ([`HOMER`](http://homer.ucsd.edu/homer/download.html)) 6. Create consensus peakset across all samples and create tabular file to aid in the filtering of the data ([`BEDTools`](https://github.com/arq5x/bedtools2/)) From 2e8c9796e88930c1bb34ac34294ea2c35c0b5cee Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sun, 15 Mar 2020 19:02:46 +0000 Subject: [PATCH 134/226] Fix section headers --- main.nf | 113 +++++++++++++++++++++++++++----------------------------- 1 file changed, 54 insertions(+), 59 deletions(-) diff --git a/main.nf b/main.nf index 0bcbcee2..05f716af 100644 --- a/main.nf +++ b/main.nf @@ -100,25 +100,28 @@ def helpMessage() { /////////////////////////////////////////////////////////////////////////////// /////////////////////////////////////////////////////////////////////////////// -/* - * SET UP CONFIGURATION VARIABLES - */ - // Show help message if (params.help) { helpMessage() exit 0 } -// Check if genome exists in the config file -if (params.genomes && params.genome && !params.genomes.containsKey(params.genome)) { - exit 1, "The provided genome '${params.genome}' is not available in the iGenomes file. Currently the available genomes are ${params.genomes.keySet().join(", ")}" +// Has the run name been specified by the user? +// this has the bonus effect of catching both -name and --name +custom_runName = params.name +if (!(workflow.runName ==~ /[a-z]+_[a-z]+/)) { + custom_runName = workflow.runName } //////////////////////////////////////////////////// /* -- DEFAULT PARAMETER VALUES -- */ //////////////////////////////////////////////////// +// Check if genome exists in the config file +if (params.genomes && params.genome && !params.genomes.containsKey(params.genome)) { + exit 1, "The provided genome '${params.genome}' is not available in the iGenomes file. Currently the available genomes are ${params.genomes.keySet().join(", ")}" +} + // Configurable variables params.fasta = params.genome ? params.genomes[ params.genome ].fasta ?: false : false params.bwa_index = params.genome ? params.genomes[ params.genome ].bwa ?: false : false @@ -132,13 +135,6 @@ params.anno_readme = params.genome ? params.genomes[ params.genome ].readme ?: f // Global variables def PEAK_TYPE = params.narrow_peak ? "narrowPeak" : "broadPeak" -// Has the run name been specified by the user? -// this has the bonus effect of catching both -name and --name -custom_runName = params.name -if (!(workflow.runName ==~ /[a-z]+_[a-z]+/)) { - custom_runName = workflow.runName -} - //////////////////////////////////////////////////// /* -- CONFIG FILES -- */ //////////////////////////////////////////////////// @@ -172,19 +168,12 @@ ch_mrep_deseq2_clustering_header = file("$baseDir/assets/multiqc/mrep_deseq2_clu /* -- VALIDATE INPUTS -- */ //////////////////////////////////////////////////// -// Validate inputs if (params.input) { ch_input = file(params.input, checkIfExists: true) } else { exit 1, "Samples design file not specified!" } if (params.gtf) { ch_gtf = file(params.gtf, checkIfExists: true) } else { exit 1, "GTF annotation file not specified!" } if (params.gene_bed) { ch_gene_bed = file(params.gene_bed, checkIfExists: true) } if (params.tss_bed) { ch_tss_bed = file(params.tss_bed, checkIfExists: true) } if (params.blacklist) { ch_blacklist = Channel.fromPath(params.blacklist, checkIfExists: true) } else { ch_blacklist = Channel.empty() } -// Save AWS IGenomes file conatining annotation version -if (params.anno_readme && file(params.anno_readme).exists()) { - file("${params.outdir}/genome/").mkdirs() - file(params.anno_readme).copyTo("${params.outdir}/genome/") -} - if (params.fasta) { lastPath = params.fasta.lastIndexOf(File.separator) bwa_base = params.fasta.substring(lastPath+1) @@ -202,6 +191,12 @@ if (params.bwa_index) { .set { ch_bwa_index } } +// Save AWS IGenomes file containing annotation version +if (params.anno_readme && file(params.anno_readme).exists()) { + file("${params.outdir}/genome/").mkdirs() + file(params.anno_readme).copyTo("${params.outdir}/genome/") +} + //////////////////////////////////////////////////// /* -- AWS -- */ //////////////////////////////////////////////////// @@ -319,7 +314,7 @@ if (!params.macs_gsize) { /////////////////////////////////////////////////////////////////////////////// /* - * PREPROCESSING - REFORMAT DESIGN FILE AND CHECK VALIDITY + * PREPROCESSING: Reformat design file and check validitiy */ process CheckDesign { tag "$design" @@ -382,7 +377,7 @@ multipleGroups = design_multiple_samples /////////////////////////////////////////////////////////////////////////////// /* - * PREPROCESSING - Build BWA index + * PREPROCESSING: Build BWA index */ if (!params.bwa_index) { process BWAIndex { @@ -406,7 +401,7 @@ if (!params.bwa_index) { } /* - * PREPROCESSING - Generate gene BED file + * PREPROCESSING: Generate gene BED file */ if (!params.gene_bed) { process MakeGeneBED { @@ -428,7 +423,7 @@ if (!params.gene_bed) { } /* - * PREPROCESSING - Generate TSS BED file + * PREPROCESSING: Generate TSS BED file */ if (!params.tss_bed) { process MakeTSSBED { @@ -449,7 +444,7 @@ if (!params.tss_bed) { } /* - * PREPROCESSING - Prepare genome intervals for filtering + * PREPROCESSING: Prepare genome intervals for filtering */ process MakeGenomeFilter { tag "$fasta" @@ -488,7 +483,7 @@ process MakeGenomeFilter { /////////////////////////////////////////////////////////////////////////////// /* - * STEP 1 - FastQC + * STEP 1: FastQC */ process FastQC { tag "$name" @@ -533,7 +528,7 @@ process FastQC { /////////////////////////////////////////////////////////////////////////////// /* - * STEP 2 - Trim Galore! + * STEP 2: Trim Galore! */ if (params.skip_trimming) { ch_trimmed_reads = ch_raw_reads_trimgalore @@ -603,7 +598,7 @@ if (params.skip_trimming) { /////////////////////////////////////////////////////////////////////////////// /* - * STEP 3.1 - Align read 1 with bwa + * STEP 3.1: Map read(s) with bwa mem */ process BWAMem { tag "$name" @@ -634,7 +629,7 @@ process BWAMem { } /* - * STEP 3.2 - Convert .bam to coordinate sorted .bam + * STEP 3.2: Convert BAM to coordinate sorted BAM */ process SortBAM { tag "$name" @@ -676,7 +671,7 @@ process SortBAM { /////////////////////////////////////////////////////////////////////////////// /* - * STEP 4.1 Merge BAM files for all libraries from same replicate + * STEP 4.1: Merge BAM files for all libraries from same sample replicate */ ch_sort_bam_merge .map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] } @@ -759,7 +754,7 @@ process MergedLibBAM { } /* - * STEP 4.2 Filter BAM file at merged library-level + * STEP 4.2: Filter BAM file at merged library-level */ process MergedLibBAMFilter { tag "$name" @@ -814,7 +809,7 @@ process MergedLibBAMFilter { } /* - * STEP 4.3 Remove orphan reads from paired-end BAM file + * STEP 4.3: Remove orphan reads from paired-end BAM file */ if (params.single_end) { ch_mlib_filter_bam @@ -886,7 +881,7 @@ if (params.single_end) { /////////////////////////////////////////////////////////////////////////////// /* - * STEP 5.1 preseq analysis after merging libraries and before filtering + * STEP 5.1: Preseq analysis after merging libraries and before filtering */ process MergedLibPreseq { tag "$name" @@ -910,7 +905,7 @@ process MergedLibPreseq { } /* - * STEP 5.2 Picard CollectMultipleMetrics after merging libraries and filtering + * STEP 5.2: Picard CollectMultipleMetrics after merging libraries and filtering */ process MergedLibMetrics { tag "$name" @@ -952,7 +947,7 @@ process MergedLibMetrics { } /* - * STEP 5.3 Read depth normalised bigWig + * STEP 5.3: Read depth normalised bigWig */ process MergedLibBigWig { tag "$name" @@ -989,7 +984,7 @@ process MergedLibBigWig { } /* - * STEP 5.4 generate gene body coverage plot with deepTools + * STEP 5.4: Generate gene body coverage plot with deepTools plotProfile */ process MergedLibPlotProfile { tag "$name" @@ -1029,7 +1024,7 @@ process MergedLibPlotProfile { } /* - * STEP 5.5 deepTools plotFingerprint + * STEP 5.5: deepTools plotFingerprint */ process MergedLibPlotFingerprint { tag "$name" @@ -1072,7 +1067,7 @@ process MergedLibPlotFingerprint { /////////////////////////////////////////////////////////////////////////////// /* - * STEP 6.1 Call peaks with MACS2 and calculate FRiP score + * STEP 6.1: Call peaks with MACS2 and calculate FRiP score */ process MergedLibMACSCallPeak { tag "$name" @@ -1127,7 +1122,7 @@ process MergedLibMACSCallPeak { } /* - * STEP 6.2 Annotate peaks with HOMER + * STEP 6.2: Annotate peaks with HOMER */ process MergedLibAnnotatePeaks { tag "$name" @@ -1159,7 +1154,7 @@ process MergedLibAnnotatePeaks { } /* - * STEP 6.3 Aggregated QC plots for peaks, FRiP and peak-to-gene annotation + * STEP 6.3: Aggregated QC plots for peaks, FRiP and peak-to-gene annotation */ process MergedLibPeakQC { label "process_medium" @@ -1197,7 +1192,7 @@ process MergedLibPeakQC { } /* - * STEP 6.4 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection + * STEP 6.4: Consensus peaks across samples, create boolean filtering file, SAF file for featureCounts and UpSetR plot for intersection */ process MergedLibConsensusPeakSet { label 'process_long' @@ -1250,7 +1245,7 @@ process MergedLibConsensusPeakSet { } /* - * STEP 6.5 Annotate consensus peaks with HOMER, and add annotation to boolean output file + * STEP 6.5: Annotate consensus peaks with HOMER, and add annotation to boolean output file */ process MergedLibConsensusPeakSetAnnotate { label "process_medium" @@ -1285,7 +1280,7 @@ process MergedLibConsensusPeakSetAnnotate { } /* - * STEP 6.6 Count reads in consensus peaks with featureCounts + * STEP 6.6: Count reads in consensus peaks with featureCounts */ process MergedLibConsensusPeakSetCounts { label 'process_medium' @@ -1320,7 +1315,7 @@ process MergedLibConsensusPeakSetCounts { } /* - * STEP 6.7 Perform differential analysis with DESeq2 + * STEP 6.7: Differential analysis with DESeq2 */ process MergedLibConsensusPeakSetDESeq { label 'process_medium' @@ -1368,7 +1363,7 @@ process MergedLibConsensusPeakSetDESeq { } /* - * STEP 6.8 Run ataqv on BAM file and corresponding peaks + * STEP 6.8: Run ataqv on BAM file and corresponding peaks */ process MergedLibAtaqv { tag "$name" @@ -1406,7 +1401,7 @@ process MergedLibAtaqv { } /* - * STEP 6.9 run ataqv mkarv on all JSON files to render web app + * STEP 6.9: Run ataqv mkarv on all JSON files to render web app */ process MergedLibAtaqvMkarv { label 'process_medium' @@ -1440,7 +1435,7 @@ process MergedLibAtaqvMkarv { /////////////////////////////////////////////////////////////////////////////// /* - * STEP 7 Merge library BAM files across all replicates + * STEP 7: Merge library BAM files across all replicates */ ch_mlib_rm_orphan_bam_mrep .map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] } @@ -1529,7 +1524,7 @@ process MergedRepBAM { /////////////////////////////////////////////////////////////////////////////// /* - * STEP 8.1 Read depth normalised bigWig + * STEP 8.1: Read depth normalised bigWig */ process MergedRepBigWig { tag "$name" @@ -1569,7 +1564,7 @@ process MergedRepBigWig { } /* - * STEP 8.2 Call peaks with MACS2 and calculate FRiP score + * STEP 8.2: Call peaks with MACS2 and calculate FRiP score */ process MergedRepMACSCallPeak { tag "$name" @@ -1623,7 +1618,7 @@ process MergedRepMACSCallPeak { } /* - * STEP 8.3 Annotate peaks with HOMER + * STEP 8.3: Annotate peaks with HOMER */ process MergedRepAnnotatePeaks { tag "$name" @@ -1655,7 +1650,7 @@ process MergedRepAnnotatePeaks { } /* - * STEP 8.4 Aggregated QC plots for peaks, FRiP and peak-to-gene annotation + * STEP 8.4: Aggregated QC plots for peaks, FRiP and peak-to-gene annotation */ process MergedRepPeakQC { label "process_medium" @@ -1693,7 +1688,7 @@ process MergedRepPeakQC { } /* - * STEP 8.5 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection + * STEP 8.5: Consensus peaks across samples, create boolean filtering file, SAF file for featureCounts and UpSetR plot for intersection */ process MergedRepConsensusPeakSet { label 'process_long' @@ -1745,7 +1740,7 @@ process MergedRepConsensusPeakSet { } /* - * STEP 8.6 Annotate consensus peaks with HOMER, and add annotation to boolean output file + * STEP 8.6: Annotate consensus peaks with HOMER, and add annotation to boolean output file */ process MergedRepConsensusPeakSetAnnotate { label "process_medium" @@ -1780,7 +1775,7 @@ process MergedRepConsensusPeakSetAnnotate { } /* - * STEP 8.7 Count reads in consensus peaks with featureCounts + * STEP 8.7: Count reads in consensus peaks with featureCounts */ process MergedRepConsensusPeakSetCounts { label 'process_medium' @@ -1815,7 +1810,7 @@ process MergedRepConsensusPeakSetCounts { } /* - * STEP 8.8 Perform differential analysis with DESeq2 + * STEP 8.8: Differential analysis with DESeq2 */ process MergedRepConsensusPeakSetDESeq { label 'process_medium' @@ -1871,7 +1866,7 @@ process MergedRepConsensusPeakSetDESeq { /////////////////////////////////////////////////////////////////////////////// /* - * STEP 9 - Create IGV session file + * STEP 9: Create IGV session file */ process IGV { publishDir "${params.outdir}/igv/${PEAK_TYPE}", mode: 'copy' @@ -1965,7 +1960,7 @@ Channel.from(summary.collect{ [it.key, it.value] }) .set { ch_workflow_summary } /* - * STEP 10 - MultiQC + * STEP 10: MultiQC */ process MultiQC { publishDir "${params.outdir}/multiqc/${PEAK_TYPE}", mode: 'copy' @@ -2031,7 +2026,7 @@ process MultiQC { /////////////////////////////////////////////////////////////////////////////// /* - * STEP 11 - Output description HTML + * STEP 11: Output description HTML */ process output_documentation { publishDir "${params.outdir}/Documentation", mode: 'copy' From 5011d65af02a674efccc583857af40ce7b352129 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sun, 15 Mar 2020 19:37:50 +0000 Subject: [PATCH 135/226] Update CHANGELOG --- CHANGELOG.md | 1 + 1 file changed, 1 insertion(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index 5d65c7eb..07410391 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -17,6 +17,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * Parameters: * `--skip_consensus_peaks` to skip consensus peak generation * `--deseq2_vst` to use variance stabilizing transformation (VST) instead of regularized log transformation (rlog) with DESeq2 + * `--publish_dir_mode` to customise method of publishing results to output directory [nf-core/tools#585](https://github.com/nf-core/tools/issues/585) ### `Fixed` From f206945ea413f57184e6bff454fe4f980dc22365 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sun, 15 Mar 2020 19:37:58 +0000 Subject: [PATCH 136/226] Add docs for --publish_dir_mode --- docs/usage.md | 5 +++++ 1 file changed, 5 insertions(+) diff --git a/docs/usage.md b/docs/usage.md index 21ed1171..c5f0d7a8 100755 --- a/docs/usage.md +++ b/docs/usage.md @@ -55,6 +55,7 @@ * [`--awscli`](#--awscli) * [Other command line parameters](#other-command-line-parameters) * [`--outdir`](#--outdir) + * [`--publish_dir_mode`](#--publish_dir_mode) * [`--email`](#--email) * [`--email_on_fail`](#--email_on_fail) * [`--max_multiqc_email_size`](#--max_multiqc_email_size) @@ -465,6 +466,10 @@ Please make sure to also set the `-w/--work-dir` and `--outdir` parameters to a The output directory where the results will be saved. +### `--publish_dir_mode` + +Value passed to Nextflow [`publishDir`](https://www.nextflow.io/docs/latest/process.html#publishdir) directive for publishing results in the output directory. Available: 'symlink', 'rellink', 'link', 'copy', 'copyNoFollow' and 'move' (Default: 'copy'). + ### `--email` Set this parameter to your e-mail address to get a summary e-mail with details of the run sent to you when the workflow exits. If set in your user config file (`~/.nextflow/config`) then you don't need to specify this on the command line for every run. From 71214275297a5c5362a4ee225f5234d5d45fdd75 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sun, 15 Mar 2020 19:38:10 +0000 Subject: [PATCH 137/226] Add --publish_dir_mode --- nextflow.config | 1 + 1 file changed, 1 insertion(+) diff --git a/nextflow.config b/nextflow.config index 97d2c64c..f13b839f 100755 --- a/nextflow.config +++ b/nextflow.config @@ -71,6 +71,7 @@ params { // Options: Other help = false outdir = './results' + publish_dir_mode = 'copy' igenomes_base = 's3://ngi-igenomes/igenomes/' igenomes_ignore = false max_multiqc_email_size = 25.MB From 2d924fdf457ec7c2a9b7407c983b6f897ce2ae80 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sun, 15 Mar 2020 19:38:24 +0000 Subject: [PATCH 138/226] Replace copy with --publish_dir_mode --- main.nf | 77 +++++++++++++++++++++++++++++---------------------------- 1 file changed, 39 insertions(+), 38 deletions(-) diff --git a/main.nf b/main.nf index 05f716af..a801fcdd 100644 --- a/main.nf +++ b/main.nf @@ -80,6 +80,7 @@ def helpMessage() { Other --outdir [file] The output directory where the results will be saved + --publish_dir_mode [str] Mode for publishing results in the output directory. Available: symlink, rellink, link, copy, copyNoFollow, move (Default: copy) --email [email] Set this parameter to your e-mail address to get a summary e-mail with details of the run sent to you when the workflow exits --email_on_fail [email] Same as --email, except only send mail if the workflow is not successful --max_multiqc_email_size [str] Theshold size for MultiQC report to be attached in notification email. If file generated by pipeline exceeds the threshold, it will not be attached (Default: 25MB) @@ -318,7 +319,7 @@ if (!params.macs_gsize) { */ process CheckDesign { tag "$design" - publishDir "${params.outdir}/pipeline_info", mode: 'copy' + publishDir "${params.outdir}/pipeline_info", mode: params.publish_dir_mode input: file design from ch_input @@ -384,7 +385,7 @@ if (!params.bwa_index) { tag "$fasta" label 'process_high' publishDir path: { params.save_reference ? "${params.outdir}/genome" : params.outdir }, - saveAs: { params.save_reference ? it : null }, mode: 'copy' + saveAs: { params.save_reference ? it : null }, mode: params.publish_dir_mode input: file fasta from ch_fasta @@ -407,7 +408,7 @@ if (!params.gene_bed) { process MakeGeneBED { tag "$gtf" label 'process_low' - publishDir "${params.outdir}/genome", mode: 'copy' + publishDir "${params.outdir}/genome", mode: params.publish_dir_mode input: file gtf from ch_gtf @@ -428,7 +429,7 @@ if (!params.gene_bed) { if (!params.tss_bed) { process MakeTSSBED { tag "$bed" - publishDir "${params.outdir}/genome", mode: 'copy' + publishDir "${params.outdir}/genome", mode: params.publish_dir_mode input: file bed from ch_gene_bed @@ -448,7 +449,7 @@ if (!params.tss_bed) { */ process MakeGenomeFilter { tag "$fasta" - publishDir "${params.outdir}/genome", mode: 'copy' + publishDir "${params.outdir}/genome", mode: params.publish_dir_mode input: file fasta from ch_fasta @@ -488,7 +489,7 @@ process MakeGenomeFilter { process FastQC { tag "$name" label 'process_medium' - publishDir "${params.outdir}/fastqc", mode: 'copy', + publishDir "${params.outdir}/fastqc", mode: params.publish_dir_mode, saveAs: { filename -> filename.endsWith(".zip") ? "zips/$filename" : "$filename" } @@ -538,7 +539,7 @@ if (params.skip_trimming) { process TrimGalore { tag "$name" label 'process_high' - publishDir "${params.outdir}/trim_galore", mode: 'copy', + publishDir "${params.outdir}/trim_galore", mode: params.publish_dir_mode, saveAs: { filename -> if (filename.endsWith(".html")) "fastqc/$filename" else if (filename.endsWith(".zip")) "fastqc/zips/$filename" @@ -635,7 +636,7 @@ process SortBAM { tag "$name" label 'process_medium' if (params.save_align_intermeds) { - publishDir path: "${params.outdir}/bwa/library", mode: 'copy', + publishDir path: "${params.outdir}/bwa/library", mode: params.publish_dir_mode, saveAs: { filename -> if (filename.endsWith(".flagstat")) "samtools_stats/$filename" else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" @@ -682,7 +683,7 @@ ch_sort_bam_merge process MergedLibBAM { tag "$name" label 'process_medium' - publishDir "${params.outdir}/bwa/mergedLibrary", mode: 'copy', + publishDir "${params.outdir}/bwa/mergedLibrary", mode: params.publish_dir_mode, saveAs: { filename -> if (filename.endsWith(".flagstat")) "samtools_stats/$filename" else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" @@ -759,7 +760,7 @@ process MergedLibBAM { process MergedLibBAMFilter { tag "$name" label 'process_medium' - publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', + publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: params.publish_dir_mode, saveAs: { filename -> if (params.single_end || params.save_align_intermeds) { if (filename.endsWith(".flagstat")) "samtools_stats/$filename" @@ -832,7 +833,7 @@ if (params.single_end) { process MergedLibBAMRemoveOrphan { tag "$name" label 'process_medium' - publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', + publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: params.publish_dir_mode, saveAs: { filename -> if (filename.endsWith(".flagstat")) "samtools_stats/$filename" else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" @@ -886,7 +887,7 @@ if (params.single_end) { process MergedLibPreseq { tag "$name" label 'process_low' - publishDir "${params.outdir}/bwa/mergedLibrary/preseq", mode: 'copy' + publishDir "${params.outdir}/bwa/mergedLibrary/preseq", mode: params.publish_dir_mode when: !params.skip_preseq @@ -910,7 +911,7 @@ process MergedLibPreseq { process MergedLibMetrics { tag "$name" label 'process_medium' - publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', + publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: params.publish_dir_mode, saveAs: { filename -> if (filename.endsWith("_metrics")) "picard_metrics/$filename" else if (filename.endsWith(".pdf")) "picard_metrics/pdf/$filename" @@ -952,7 +953,7 @@ process MergedLibMetrics { process MergedLibBigWig { tag "$name" label 'process_medium' - publishDir "${params.outdir}/bwa/mergedLibrary/bigwig", mode: 'copy', + publishDir "${params.outdir}/bwa/mergedLibrary/bigwig", mode: params.publish_dir_mode, saveAs: { filename -> if (filename.endsWith("scale_factor.txt")) "scale/$filename" else if (filename.endsWith(".bigWig")) "$filename" @@ -989,7 +990,7 @@ process MergedLibBigWig { process MergedLibPlotProfile { tag "$name" label 'process_high' - publishDir "${params.outdir}/bwa/mergedLibrary/deepTools/plotProfile", mode: 'copy' + publishDir "${params.outdir}/bwa/mergedLibrary/deepTools/plotProfile", mode: params.publish_dir_mode when: !params.skip_plot_profile @@ -1029,7 +1030,7 @@ process MergedLibPlotProfile { process MergedLibPlotFingerprint { tag "$name" label 'process_high' - publishDir "${params.outdir}/bwa/mergedLibrary/deepTools/plotFingerprint", mode: 'copy' + publishDir "${params.outdir}/bwa/mergedLibrary/deepTools/plotFingerprint", mode: params.publish_dir_mode when: !params.skip_plot_fingerprint @@ -1072,7 +1073,7 @@ process MergedLibPlotFingerprint { process MergedLibMACSCallPeak { tag "$name" label 'process_medium' - publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}", mode: 'copy', + publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}", mode: params.publish_dir_mode, saveAs: { filename -> if (filename.endsWith(".tsv")) "qc/$filename" else if (filename.endsWith(".igv.txt")) null @@ -1127,7 +1128,7 @@ process MergedLibMACSCallPeak { process MergedLibAnnotatePeaks { tag "$name" label "process_medium" - publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}", mode: 'copy' + publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}", mode: params.publish_dir_mode when: params.macs_gsize @@ -1158,7 +1159,7 @@ process MergedLibAnnotatePeaks { */ process MergedLibPeakQC { label "process_medium" - publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/qc", mode: 'copy' + publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/qc", mode: params.publish_dir_mode when: params.macs_gsize @@ -1196,7 +1197,7 @@ process MergedLibPeakQC { */ process MergedLibConsensusPeakSet { label 'process_long' - publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: 'copy', + publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: params.publish_dir_mode, saveAs: { filename -> if (filename.endsWith(".igv.txt")) null else filename @@ -1249,7 +1250,7 @@ process MergedLibConsensusPeakSet { */ process MergedLibConsensusPeakSetAnnotate { label "process_medium" - publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: 'copy' + publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: params.publish_dir_mode when: params.macs_gsize && (replicatesExist || multipleGroups) && !params.skip_consensus_peaks @@ -1284,7 +1285,7 @@ process MergedLibConsensusPeakSetAnnotate { */ process MergedLibConsensusPeakSetCounts { label 'process_medium' - publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: 'copy' + publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: params.publish_dir_mode when: params.macs_gsize && (replicatesExist || multipleGroups) && !params.skip_consensus_peaks @@ -1319,7 +1320,7 @@ process MergedLibConsensusPeakSetCounts { */ process MergedLibConsensusPeakSetDESeq { label 'process_medium' - publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus/deseq2", mode: 'copy', + publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus/deseq2", mode: params.publish_dir_mode, saveAs: { filename -> if (filename.endsWith(".igv.txt")) null else filename @@ -1368,7 +1369,7 @@ process MergedLibConsensusPeakSetDESeq { process MergedLibAtaqv { tag "$name" label 'process_medium' - publishDir "${params.outdir}/bwa/mergedLibrary/ataqv/${PEAK_TYPE}", mode: 'copy' + publishDir "${params.outdir}/bwa/mergedLibrary/ataqv/${PEAK_TYPE}", mode: params.publish_dir_mode when: !params.skip_ataqv @@ -1405,7 +1406,7 @@ process MergedLibAtaqv { */ process MergedLibAtaqvMkarv { label 'process_medium' - publishDir "${params.outdir}/bwa/mergedLibrary/ataqv/${PEAK_TYPE}", mode: 'copy' + publishDir "${params.outdir}/bwa/mergedLibrary/ataqv/${PEAK_TYPE}", mode: params.publish_dir_mode when: !params.skip_ataqv @@ -1446,7 +1447,7 @@ ch_mlib_rm_orphan_bam_mrep process MergedRepBAM { tag "$name" label 'process_medium' - publishDir "${params.outdir}/bwa/mergedReplicate", mode: 'copy', + publishDir "${params.outdir}/bwa/mergedReplicate", mode: params.publish_dir_mode, saveAs: { filename -> if (filename.endsWith(".flagstat")) "samtools_stats/$filename" else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" @@ -1529,7 +1530,7 @@ process MergedRepBAM { process MergedRepBigWig { tag "$name" label 'process_medium' - publishDir "${params.outdir}/bwa/mergedReplicate/bigwig", mode: 'copy', + publishDir "${params.outdir}/bwa/mergedReplicate/bigwig", mode: params.publish_dir_mode, saveAs: { filename -> if (filename.endsWith("scale_factor.txt")) "scale/$filename" else if (filename.endsWith(".bigWig")) "$filename" @@ -1569,7 +1570,7 @@ process MergedRepBigWig { process MergedRepMACSCallPeak { tag "$name" label 'process_medium' - publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}", mode: 'copy', + publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}", mode: params.publish_dir_mode, saveAs: { filename -> if (filename.endsWith(".tsv")) "qc/$filename" else if (filename.endsWith(".igv.txt")) null @@ -1623,7 +1624,7 @@ process MergedRepMACSCallPeak { process MergedRepAnnotatePeaks { tag "$name" label "process_medium" - publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}", mode: 'copy' + publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}", mode: params.publish_dir_mode when: !params.skip_merge_replicates && replicatesExist && params.macs_gsize @@ -1654,7 +1655,7 @@ process MergedRepAnnotatePeaks { */ process MergedRepPeakQC { label "process_medium" - publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/qc", mode: 'copy' + publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/qc", mode: params.publish_dir_mode when: !params.skip_merge_replicates && replicatesExist && params.macs_gsize @@ -1692,7 +1693,7 @@ process MergedRepPeakQC { */ process MergedRepConsensusPeakSet { label 'process_long' - publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: 'copy', + publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: params.publish_dir_mode, saveAs: { filename -> if (filename.endsWith(".igv.txt")) null else filename @@ -1744,7 +1745,7 @@ process MergedRepConsensusPeakSet { */ process MergedRepConsensusPeakSetAnnotate { label "process_medium" - publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: 'copy' + publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: params.publish_dir_mode when: !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks @@ -1779,7 +1780,7 @@ process MergedRepConsensusPeakSetAnnotate { */ process MergedRepConsensusPeakSetCounts { label 'process_medium' - publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: 'copy' + publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: params.publish_dir_mode when: !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks @@ -1814,7 +1815,7 @@ process MergedRepConsensusPeakSetCounts { */ process MergedRepConsensusPeakSetDESeq { label 'process_medium' - publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus/deseq2", mode: 'copy', + publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus/deseq2", mode: params.publish_dir_mode, saveAs: { filename -> if (filename.endsWith(".igv.txt")) null else filename @@ -1869,7 +1870,7 @@ process MergedRepConsensusPeakSetDESeq { * STEP 9: Create IGV session file */ process IGV { - publishDir "${params.outdir}/igv/${PEAK_TYPE}", mode: 'copy' + publishDir "${params.outdir}/igv/${PEAK_TYPE}", mode: params.publish_dir_mode when: !params.skip_igv @@ -1909,7 +1910,7 @@ process IGV { * Parse software version numbers */ process get_software_versions { - publishDir "${params.outdir}/pipeline_info", mode: 'copy', + publishDir "${params.outdir}/pipeline_info", mode: params.publish_dir_mode, saveAs: { filename -> if (filename.indexOf(".csv") > 0) filename else null @@ -1963,7 +1964,7 @@ Channel.from(summary.collect{ [it.key, it.value] }) * STEP 10: MultiQC */ process MultiQC { - publishDir "${params.outdir}/multiqc/${PEAK_TYPE}", mode: 'copy' + publishDir "${params.outdir}/multiqc/${PEAK_TYPE}", mode: params.publish_dir_mode when: !params.skip_multiqc @@ -2029,7 +2030,7 @@ process MultiQC { * STEP 11: Output description HTML */ process output_documentation { - publishDir "${params.outdir}/Documentation", mode: 'copy' + publishDir "${params.outdir}/Documentation", mode: params.publish_dir_mode input: file output_docs from ch_output_docs From 4fe6230a7bdb0f2e157487b6cb19326d606c1cd8 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Mon, 16 Mar 2020 11:49:46 +0000 Subject: [PATCH 139/226] Units of 6GB for GCP --- conf/base.config | 8 ++++---- 1 file changed, 4 insertions(+), 4 deletions(-) diff --git a/conf/base.config b/conf/base.config index f1a6373c..92a48f32 100755 --- a/conf/base.config +++ b/conf/base.config @@ -12,7 +12,7 @@ process { cpus = { check_max( 1 * task.attempt, 'cpus' ) } - memory = { check_max( 7.GB * task.attempt, 'memory' ) } + memory = { check_max( 6.GB * task.attempt, 'memory' ) } time = { check_max( 4.h * task.attempt, 'time' ) } errorStrategy = { task.exitStatus in [143,137,104,134,139] ? 'retry' : 'finish' } @@ -22,17 +22,17 @@ process { // Process-specific resource requirements withLabel:process_low { cpus = { check_max( 2 * task.attempt, 'cpus' ) } - memory = { check_max( 14.GB * task.attempt, 'memory' ) } + memory = { check_max( 12.GB * task.attempt, 'memory' ) } time = { check_max( 6.h * task.attempt, 'time' ) } } withLabel:process_medium { cpus = { check_max( 6 * task.attempt, 'cpus' ) } - memory = { check_max( 42.GB * task.attempt, 'memory' ) } + memory = { check_max( 36.GB * task.attempt, 'memory' ) } time = { check_max( 8.h * task.attempt, 'time' ) } } withLabel:process_high { cpus = { check_max( 12 * task.attempt, 'cpus' ) } - memory = { check_max( 84.GB * task.attempt, 'memory' ) } + memory = { check_max( 72.GB * task.attempt, 'memory' ) } time = { check_max( 16.h * task.attempt, 'time' ) } } withLabel:process_long { From 1933d43e3b9bc2ac6cc7e7f0e47395a63932df28 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Mon, 16 Mar 2020 12:07:44 +0000 Subject: [PATCH 140/226] Add matrix tests --- .github/workflows/ci.yml | 22 ++++++++++++++++++++++ 1 file changed, 22 insertions(+) diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml index 2f69d7c5..a3cff843 100644 --- a/.github/workflows/ci.yml +++ b/.github/workflows/ci.yml @@ -26,3 +26,25 @@ jobs: - name: Run pipeline with test data run: | nextflow run ${GITHUB_WORKSPACE} -profile test,docker + + parameters: + env: + NXF_VER: '19.10.0' + NXF_ANSI_LOG: false + runs-on: ubuntu-latest + strategy: + matrix: + parameters: [--single_end, --skip_trimming, --skip_merge_replicates, --skip_consensus_peaks, --skip_multiqc] + steps: + - uses: actions/checkout@v2 + - name: Install Nextflow + run: | + wget -qO- get.nextflow.io | bash + sudo mv nextflow /usr/local/bin/ + - name: Pull docker image + run: | + docker pull nfcore/atacseq:dev + docker tag nfcore/atacseq:dev nfcore/atacseq:1.2.0 + - name: Run pipeline with various options + run: | + nextflow run ${GITHUB_WORKSPACE} -profile test,docker ${{ matrix.parameters }} From 1bce248549a7f843fbc53b280fcd495e2bd4a908 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 17 Mar 2020 16:41:05 +0000 Subject: [PATCH 141/226] Fix tyop --- main.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/main.nf b/main.nf index a801fcdd..185138b4 100644 --- a/main.nf +++ b/main.nf @@ -45,7 +45,7 @@ def helpMessage() { --clip_r1 [int] Instructs Trim Galore to remove bp from the 5' end of read 1 (or single-end reads) (Default: 0) --clip_r2 [int] Instructs Trim Galore to remove bp from the 5' end of read 2 (paired-end reads only) (Default: 0) --three_prime_clip_r1 [int] Instructs Trim Galore to remove bp from the 3' end of read 1 AFTER adapter/quality trimming has been performed (Default: 0) - --three_prime_clip_r2 [int] Instructs Trim Galore to re move bp from the 3' end of read 2 AFTER adapter/quality trimming has been performed (Default: 0) + --three_prime_clip_r2 [int] Instructs Trim Galore to remove bp from the 3' end of read 2 AFTER adapter/quality trimming has been performed (Default: 0) --trim_nextseq [int] Instructs Trim Galore to apply the --nextseq=X option, to trim based on quality after removing poly-G tails (Default: 0) --skip_trimming [bool] Skip the adapter trimming step --save_trimmed [bool] Save the trimmed FastQ files in the results directory From 284ae30f86b1ea1658901bde1e2233ae4da08a2a Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 17 Mar 2020 16:41:25 +0000 Subject: [PATCH 142/226] First stab at generating json schema file --- nextflow_schema.json | 501 +++++++++++++++++++++++++++++++++++++++++++ 1 file changed, 501 insertions(+) create mode 100644 nextflow_schema.json diff --git a/nextflow_schema.json b/nextflow_schema.json new file mode 100644 index 00000000..f447d0ff --- /dev/null +++ b/nextflow_schema.json @@ -0,0 +1,501 @@ +{ + "$schema": "http://json-schema.org/draft-07/schema", + "$id": "https://raw.githubusercontent.com/nf-core/atacseq/master/nextflow_schema.json", + "title": "nf-core/atacseq pipeline parameters", + "description": "ATACSeq peak-calling and differential analysis pipeline.", + "type": "object", + "properties": { + "Generic options": { + "type": "object", + "description": "", + "default": "", + "properties": { + "input": { + "type": "string", + "default": "'./design.csv'", + "description": "Comma-separated file containing information about the samples in the experiment", + "help_text": "You will need to create a design file with information about the samples in your experiment before running the pipeline. Use this parameter to specify its location. It has to be a comma-separated file with 4 columns, and a header row. See [usage docs](https://nf-co.re/atacseq/docs/usage#--input)." + }, + "single_end": { + "type": "boolean", + "description": "Specifies that the input is single-end reads", + "fa_icon": "", + "default": "False", + "help_text": "By default, the pipeline expects paired-end data. If you have single-end data, specify `--single_end` on the command line when you launch the pipeline. It is not possible to run a mixture of single-end and paired-end files in one run." + }, + "fragment_size": { + "type": "integer", + "default": "0", + "description": "Estimated fragment size used to extend single-end reads" + }, + "fingerprint_bins": { + "type": "integer", + "default": "500000", + "description": "Number of genomic bins to use when calculating deepTools fingerprint plot" + }, + "seq_center": { + "type": "string", + "default": "false", + "description": "Sequencing center information to be added to read group of BAM files" + } + }, + "required": [ + "input" + ] + }, + "Reference genome options": { + "type": "object", + "description": "", + "default": "", + "properties": { + "genome": { + "type": "string", + "description": "Name of iGenomes reference", + "fa_icon": "", + "help_text": "If using a reference genome configured in the pipeline using iGenomes, use this parameter to give the ID for the reference. This is then used to build the full paths for all required reference genome files. For example: `--genome GRCh38`.", + "default": "false" + }, + "fasta": { + "type": "string", + "default": "false", + "description": "Path to Fasta reference. Not mandatory when using reference in iGenomes config via --genome", + "help_text": "Full path to fasta file containing reference genome (*mandatory* if `--genome` is not specified). If you don't have a BWA index available this will be generated for you automatically. Combine with `--save_reference` to save BWA index for future runs." + }, + "gtf": { + "type": "string", + "default": "false", + "description": "Path to GTF file. Not mandatory when using reference in iGenomes config via --genome" + }, + "bwa_index": { + "type": "string", + "default": "false", + "description": "Full path to directory containing BWA index including base name i.e. /path/to/index/genome.fa" + }, + "gene_bed": { + "type": "string", + "default": "false", + "description": "Path to BED file containing gene intervals. This will be created from the GTF file if not specified." + }, + "tss_bed": { + "type": "string", + "default": "false", + "description": "Path to BED file containing transcription start sites. This will be created from the gene BED file if not specified." + }, + "macs_gsize": { + "type": "string", + "default": "false", + "description": "Effective genome size parameter required by MACS2. If using iGenomes config, values have only been provided when --genome is set as GRCh37, GRCm38, hg19, mm10, BDGP6 and WBcel235", + "help_text": "[Effective genome size](https://github.com/taoliu/MACS#-g--gsize) parameter required by MACS2. These have been provided when `--genome` is set as *GRCh37*, *GRCh38*, *GRCm38*, *WBcel235*, *BDGP6*, *R64-1-1*, *EF2*, *hg38*, *hg19* and *mm10*. For other genomes, if this parameter is not specified then the MACS2 peak-calling and differential analysis will be skipped." + }, + "blacklist": { + "type": "string", + "default": "false", + "description": "Path to blacklist regions (.BED format), used for filtering alignments", + "help_text": "If provided, alignments that overlap with the regions in this file will be filtered out (see [ENCODE blacklists](https://sites.google.com/site/anshulkundaje/projects/blacklists)). The file should be in BED format. Blacklisted regions for *GRCh37*, *GRCh38*, *GRCm38*, *hg19*, *hg38*, *mm10* are bundled with the pipeline in the [`blacklists`](../assets/blacklists/) directory, and as such will be automatically used if any of those genomes are specified with the `--genome` parameter." + }, + "mito_name": { + "type": "string", + "default": "false", + "description": "Name of Mitochondrial chomosome in genome fasta (e.g. chrM). Reads aligning to this contig are filtered out", + "help_text": "Name of mitochondrial chomosome in reference assembly. Reads aligning to this contig are filtered out if a valid identifier is provided otherwise this step is skipped. Where possible these have been provided in the [`igenomes.config`](../conf/igenomes.config)." + }, + "save_reference": { + "type": "boolean", + "default": "False", + "description": "If generated by the pipeline save the BWA index in the results directory", + "help_text": "If the BWA index is generated by the pipeline use this parameter to save it to your results folder. These can then be used for future pipeline runs, reducing processing times." + }, + "igenomes_base": { + "type": "string", + "description": "Directory / URL base for iGenomes references", + "default": "s3://ngi-igenomes/igenomes/", + "fa_icon": "", + "hidden": true + }, + "igenomes_ignore": { + "type": "boolean", + "description": "Do not load the iGenomes reference config", + "fa_icon": "", + "hidden": true, + "default": "False", + "help_text": "Do not load `igenomes.config` when running the pipeline. You may choose this option if you observe clashes between custom parameters and those supplied in `igenomes.config`." + } + }, + "required": [ + "fasta", + "gtf" + ] + }, + "Adapter trimming options": { + "type": "object", + "description": "", + "default": "", + "properties": { + "clip_r1": { + "type": "integer", + "default": "0", + "description": "Instructs Trim Galore to remove bp from the 5' end of read 1 (or single-end reads)" + }, + "clip_r2": { + "type": "integer", + "default": "0", + "description": "Instructs Trim Galore to remove bp from the 5' end of read 2 (paired-end reads only)" + }, + "three_prime_clip_r1": { + "type": "integer", + "default": "0", + "description": "Instructs Trim Galore to remove bp from the 3' end of read 1 AFTER adapter/quality trimming has been performed" + }, + "three_prime_clip_r2": { + "type": "integer", + "default": "0", + "description": "Instructs Trim Galore to remove bp from the 3' end of read 2 AFTER adapter/quality trimming has been performed" + }, + "trim_nextseq": { + "type": "integer", + "default": "0", + "description": "Instructs Trim Galore to apply the --nextseq=X option, to trim based on quality after removing poly-G tails", + "help_text": "This enables the option Cutadapt `--nextseq-trim=3'CUTOFF` option via Trim Galore, which will set a quality cutoff (that is normally given with -q instead), but qualities of G bases are ignored. This trimming is in common for the NextSeq- and NovaSeq-platforms, where basecalls without any signal are called as high-quality G bases." + }, + "skip_trimming": { + "type": "boolean", + "default": "False", + "description": "Skip the adapter trimming step", + "help_text": "Skip the adapter trimming step. Use this if your input FastQ files have already been trimmed outside of the workflow or if you're very confident that there is no adapter contamination in your data." + }, + "save_trimmed": { + "type": "boolean", + "default": "False", + "description": "Save the trimmed FastQ files in the results directory", + "help_text": "By default, trimmed FastQ files will not be saved to the results directory. Specify this flag (or set to true in your config file) to copy these files to the results directory when complete." + } + } + }, + "Alignment options": { + "type": "object", + "description": "", + "default": "", + "properties": { + "keep_mito": { + "type": "boolean", + "default": "False", + "description": "Reads mapping to mitochondrial contig are not filtered from alignments" + }, + "keep_dups": { + "type": "boolean", + "default": "False", + "description": "Duplicate reads are not filtered from alignments" + }, + "keep_multi_map": { + "type": "boolean", + "default": "False", + "description": "Reads mapping to multiple locations are not filtered from alignments" + }, + "skip_merge_replicates": { + "type": "boolean", + "default": "False", + "description": "Do not perform alignment merging and downstream analysis by merging replicates i.e. only do this by merging resequenced libraries", + "help_text": "An additional series of steps are performed by the pipeline by merging the replicates from the same experimental group. This is primarily to increase the sequencing depth in order to perform downstream analyses such as footprinting. Specifying this parameter means that these steps will not be performed." + }, + "save_align_intermeds": { + "type": "boolean", + "default": "False", + "description": "Save the intermediate BAM files from the alignment step", + "help_text": "By default, intermediate BAM files will not be saved. The final BAM files created after the appropriate filtering step are always saved to limit storage usage. Set this parameter to also save other intermediate BAM files." + } + } + }, + "Peak calling options": { + "type": "object", + "description": "", + "default": "", + "properties": { + "narrow_peak": { + "type": "boolean", + "default": "False", + "description": "Run MACS2 in narrowPeak mode", + "help_text": "MACS2 is run by default with the [`--broad`](https://github.com/taoliu/MACS#--broad) flag. Specify this flag to call peaks in narrowPeak mode." + }, + "broad_cutoff": { + "type": "number", + "default": "0.1", + "description": "Specifies broad cutoff value for MACS2. Only used when --narrow_peak isnt specified", + "help_text": "Specifies broad cut-off value for MACS2. Only used when `--narrow_peak` isnt specified." + }, + "min_reps_consensus": { + "type": "integer", + "default": "1", + "description": "Number of biological replicates required from a given condition for a peak to contribute to a consensus peak", + "help_text": "Number of biological replicates required from a given condition for a peak to contribute to a consensus peak . If you are confident you have good reproducibility amongst your replicates then you can increase the value of this parameter to create a \"reproducible\" set of consensus of peaks. For example, a value of 2 will mean peaks that have been called in at least 2 replicates will contribute to the consensus set of peaks, and as such peaks that are unique to a given replicate will be discarded." + }, + "save_macs_pileup": { + "type": "boolean", + "default": "False", + "description": "Instruct MACS2 to create bedGraph files normalised to signal per million reads" + }, + "skip_consensus_peaks": { + "type": "boolean", + "default": "False", + "description": "Skip consensus peak generation, annotation and counting" + } + } + }, + "Differential analysis options": { + "type": "object", + "description": "", + "default": "", + "properties": { + "deseq2_vst": { + "type": "boolean", + "default": "False", + "description": "Use vst transformation instead of rlog with DESeq2", + "help_text": "Use `vst` transformation instead of `rlog` with DESeq2. See [DESeq2 docs](http://bioconductor.org/packages/devel/bioc/vignettes/DESeq2/inst/doc/DESeq2.html#data-transformations-and-visualization)." + }, + "skip_diff_analysis": { + "type": "boolean", + "default": "False", + "description": "Skip differential accessibility analysis" + } + } + }, + "Process skipping options": { + "type": "object", + "description": "", + "default": "", + "properties": { + "skip_fastqc": { + "type": "boolean", + "default": "False", + "description": "Skip FastQC" + }, + "skip_picard_metrics": { + "type": "boolean", + "default": "False", + "description": "Skip Picard CollectMultipleMetrics" + }, + "skip_preseq": { + "type": "boolean", + "default": "False", + "description": "Skip Preseq" + }, + "skip_plot_profile": { + "type": "boolean", + "default": "False", + "description": "Skip deepTools plotProfile" + }, + "skip_plot_fingerprint": { + "type": "boolean", + "default": "False", + "description": "Skip deepTools plotFingerprint" + }, + "skip_ataqv": { + "type": "boolean", + "default": "False", + "description": "Skip Ataqv" + }, + "skip_igv": { + "type": "boolean", + "default": "False", + "description": "Skip IGV" + }, + "skip_multiqc": { + "type": "boolean", + "default": "False", + "description": "Skip MultiQC" + } + } + }, + "Pipeline config options": { + "type": "object", + "description": "", + "default": "", + "properties": { + "multiqc_config": { + "type": "string", + "description": "Custom config file to supply to MultiQC", + "default": "false", + "fa_icon": "", + "hidden": true + }, + "bamtools_filter_pe_config": { + "type": "string", + "default": "\"$baseDir/assets/bamtools_filter_pe.json\"", + "hidden": true, + "description": "BAMTools JSON file with custom filters for paired-end data" + }, + "bamtools_filter_se_config": { + "type": "string", + "default": "\"$baseDir/assets/bamtools_filter_se.json\"", + "hidden": true, + "description": "BAMTools JSON file with custom filters for single-end data" + } + } + }, + "Institutional config options": { + "type": "object", + "description": "", + "default": "", + "properties": { + "custom_config_version": { + "type": "string", + "description": "Git commit id for Institutional configs", + "default": "master", + "hidden": true + }, + "custom_config_base": { + "type": "string", + "description": "Base directory for Institutional configs", + "default": "https://raw.githubusercontent.com/nf-core/configs/master", + "hidden": true, + "help_text": "If you're running offline, Nextflow will not be able to fetch the institutional config files\nfrom the internet. If you don't need them, then this is not a problem. If you do need them,\nyou should download the files from the repo and tell nextflow where to find them with the `--custom_config_base` option." + }, + "hostnames": { + "type": "string", + "description": "Institutional configs hostname", + "default": "false", + "hidden": true + }, + "config_profile_description": { + "type": "string", + "description": "Institutional config description", + "hidden": true, + "default": "false" + }, + "config_profile_contact": { + "type": "string", + "description": "Institutional config contact information", + "hidden": true, + "default": "false" + }, + "config_profile_url": { + "type": "string", + "description": "Institutional config URL link", + "hidden": true, + "default": "false" + } + } + }, + "Max job request options": { + "type": "object", + "description": "", + "default": "", + "properties": { + "max_cpus": { + "type": "integer", + "description": "Maximum number of CPUs that can be requested for any single job", + "default": 16, + "fa_icon": "", + "hidden": true, + "help_text": "Use to set an upper-limit for the CPU requirement for each process. Should be an integer e.g. `--max_cpus 1`" + }, + "max_memory": { + "type": "string", + "description": "Maximum amount of memory that can be requested for any single job", + "default": "128 GB", + "fa_icon": "", + "hidden": true, + "help_text": "Use to set an upper-limit for the memory requirement for each process. Should be a string in the format integer-unit e.g. `--max_memory '8.GB'`" + }, + "max_time": { + "type": "string", + "description": "Maximum amount of time that can be requested for any single job", + "default": "10d", + "fa_icon": "", + "hidden": true, + "help_text": "Use to set an upper-limit for the time requirement for each process. Should be a string in the format integer-unit e.g. `--max_time '2.h'`" + } + } + }, + "Pipeline template options": { + "type": "object", + "description": "", + "default": "", + "properties": { + "help": { + "type": "boolean", + "description": "Display help text", + "hidden": true, + "fa_icon": "", + "default": "False" + }, + "outdir": { + "type": "string", + "description": "The output directory where the results will be saved", + "default": "./results", + "fa_icon": "" + }, + "publish_dir_mode": { + "type": "string", + "default": "copy", + "hidden": true, + "description": "Method used to save pipeline results to output directory", + "help_text": "The Nextflow `publishDir` option specifies which intermediate files should be saved to the output directory. This option tells the pipeline what method should be used to move these files. See https://www.nextflow.io/docs/latest/process.html#publishdir for details.", + "fa_icon": "", + "enum": [ + "symlink", + "rellink", + "link", + "copy", + "copyNoFollow", + "mov" + ] + }, + "name": { + "type": "string", + "description": "Workflow name", + "fa_icon": "", + "help_text": "A custom name for the pipeline run. Unlike the core nextflow `-name` option with one hyphen this parameter can be reused multiple times, for example if using `-resume`. Passed through to steps such as MultiQC and used for things like report filenames and titles.", + "default": "false" + }, + "email": { + "type": "string", + "description": "Email address for completion summary", + "fa_icon": "", + "help_text": "An email address to send a summary email to when the pipeline is completed.", + "pattern": "^([a-zA-Z0-9_\\-\\.]+)@([a-zA-Z0-9_\\-\\.]+)\\.([a-zA-Z]{2,5})$" + }, + "email_on_fail": { + "type": "string", + "description": "Email address for completion summary, only when pipeline fails", + "fa_icon": "", + "pattern": "^([a-zA-Z0-9_\\-\\.]+)@([a-zA-Z0-9_\\-\\.]+)\\.([a-zA-Z]{2,5})$", + "help_text": "An email address to send a summary email to when the pipeline is completed - ONLY sent if the pipeline does not exit successfully." + }, + "plaintext_email": { + "type": "boolean", + "description": "Send plain-text email instead of HTML", + "fa_icon": "", + "hidden": true, + "default": "False" + }, + "max_multiqc_email_size": { + "type": "string", + "description": "File size limit when attaching MultiQC reports to summary emails", + "default": "25 MB", + "fa_icon": "", + "hidden": true + }, + "monochrome_logs": { + "type": "boolean", + "description": "Do not use coloured log outputs", + "fa_icon": "", + "hidden": true, + "default": "False" + }, + "tracedir": { + "type": "string", + "description": "Directory to keep pipeline Nextflow logs and reports", + "default": "\"${params.outdir}/pipeline_info\"", + "fa_icon": "", + "hidden": true + }, + "clusterOptions": { + "type": "string", + "default": "false", + "description": "Arguments passed to Nextflow clusterOptions", + "hidden": true + } + } + } + } +} From f55f18e3586c74c82aaf66df6851f95b8cab2994 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 17 Mar 2020 17:21:36 +0000 Subject: [PATCH 143/226] Update params --- nextflow.config | 1 + 1 file changed, 1 insertion(+) diff --git a/nextflow.config b/nextflow.config index f13b839f..9e16c428 100755 --- a/nextflow.config +++ b/nextflow.config @@ -9,6 +9,7 @@ params { // Options: Generic + input = './design.csv' single_end = false seq_center = false fragment_size = 0 From f579b54e6451248c97fe9c8eebf70318cbc55fa5 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 17 Mar 2020 17:25:40 +0000 Subject: [PATCH 144/226] Update schema --- nextflow_schema.json | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/nextflow_schema.json b/nextflow_schema.json index f447d0ff..25137d7c 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -12,7 +12,7 @@ "properties": { "input": { "type": "string", - "default": "'./design.csv'", + "default": "./design.csv", "description": "Comma-separated file containing information about the samples in the experiment", "help_text": "You will need to create a design file with information about the samples in your experiment before running the pipeline. Use this parameter to specify its location. It has to be a comma-separated file with 4 columns, and a header row. See [usage docs](https://nf-co.re/atacseq/docs/usage#--input)." }, From 5e4b0a7b0570bac21622eecb2109aebbf4fd7a62 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 25 Mar 2020 12:40:07 +0000 Subject: [PATCH 145/226] Update schema --- nextflow_schema.json | 145 ++++++++++++++++--------------------------- 1 file changed, 53 insertions(+), 92 deletions(-) diff --git a/nextflow_schema.json b/nextflow_schema.json index 25137d7c..767d5691 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -7,8 +7,6 @@ "properties": { "Generic options": { "type": "object", - "description": "", - "default": "", "properties": { "input": { "type": "string", @@ -19,23 +17,22 @@ "single_end": { "type": "boolean", "description": "Specifies that the input is single-end reads", - "fa_icon": "", - "default": "False", + "fa_icon": "fas fa-align-center", + "default": false, "help_text": "By default, the pipeline expects paired-end data. If you have single-end data, specify `--single_end` on the command line when you launch the pipeline. It is not possible to run a mixture of single-end and paired-end files in one run." }, "fragment_size": { "type": "integer", - "default": "0", + "default": 0, "description": "Estimated fragment size used to extend single-end reads" }, "fingerprint_bins": { "type": "integer", - "default": "500000", + "default": 500000, "description": "Number of genomic bins to use when calculating deepTools fingerprint plot" }, "seq_center": { "type": "string", - "default": "false", "description": "Sequencing center information to be added to read group of BAM files" } }, @@ -45,63 +42,52 @@ }, "Reference genome options": { "type": "object", - "description": "", - "default": "", "properties": { "genome": { "type": "string", "description": "Name of iGenomes reference", - "fa_icon": "", + "fa_icon": "fas fa-book", "help_text": "If using a reference genome configured in the pipeline using iGenomes, use this parameter to give the ID for the reference. This is then used to build the full paths for all required reference genome files. For example: `--genome GRCh38`.", - "default": "false" }, "fasta": { "type": "string", - "default": "false", "description": "Path to Fasta reference. Not mandatory when using reference in iGenomes config via --genome", "help_text": "Full path to fasta file containing reference genome (*mandatory* if `--genome` is not specified). If you don't have a BWA index available this will be generated for you automatically. Combine with `--save_reference` to save BWA index for future runs." }, "gtf": { "type": "string", - "default": "false", "description": "Path to GTF file. Not mandatory when using reference in iGenomes config via --genome" }, "bwa_index": { "type": "string", - "default": "false", "description": "Full path to directory containing BWA index including base name i.e. /path/to/index/genome.fa" }, "gene_bed": { "type": "string", - "default": "false", "description": "Path to BED file containing gene intervals. This will be created from the GTF file if not specified." }, "tss_bed": { "type": "string", - "default": "false", "description": "Path to BED file containing transcription start sites. This will be created from the gene BED file if not specified." }, "macs_gsize": { "type": "string", - "default": "false", "description": "Effective genome size parameter required by MACS2. If using iGenomes config, values have only been provided when --genome is set as GRCh37, GRCm38, hg19, mm10, BDGP6 and WBcel235", "help_text": "[Effective genome size](https://github.com/taoliu/MACS#-g--gsize) parameter required by MACS2. These have been provided when `--genome` is set as *GRCh37*, *GRCh38*, *GRCm38*, *WBcel235*, *BDGP6*, *R64-1-1*, *EF2*, *hg38*, *hg19* and *mm10*. For other genomes, if this parameter is not specified then the MACS2 peak-calling and differential analysis will be skipped." }, "blacklist": { "type": "string", - "default": "false", "description": "Path to blacklist regions (.BED format), used for filtering alignments", "help_text": "If provided, alignments that overlap with the regions in this file will be filtered out (see [ENCODE blacklists](https://sites.google.com/site/anshulkundaje/projects/blacklists)). The file should be in BED format. Blacklisted regions for *GRCh37*, *GRCh38*, *GRCm38*, *hg19*, *hg38*, *mm10* are bundled with the pipeline in the [`blacklists`](../assets/blacklists/) directory, and as such will be automatically used if any of those genomes are specified with the `--genome` parameter." }, "mito_name": { "type": "string", - "default": "false", "description": "Name of Mitochondrial chomosome in genome fasta (e.g. chrM). Reads aligning to this contig are filtered out", "help_text": "Name of mitochondrial chomosome in reference assembly. Reads aligning to this contig are filtered out if a valid identifier is provided otherwise this step is skipped. Where possible these have been provided in the [`igenomes.config`](../conf/igenomes.config)." }, "save_reference": { "type": "boolean", - "default": "False", + "default": false, "description": "If generated by the pipeline save the BWA index in the results directory", "help_text": "If the BWA index is generated by the pipeline use this parameter to save it to your results folder. These can then be used for future pipeline runs, reducing processing times." }, @@ -109,15 +95,15 @@ "type": "string", "description": "Directory / URL base for iGenomes references", "default": "s3://ngi-igenomes/igenomes/", - "fa_icon": "", + "fa_icon": "fas fa-cloud-download-alt", "hidden": true }, "igenomes_ignore": { "type": "boolean", "description": "Do not load the iGenomes reference config", - "fa_icon": "", + "fa_icon": "fas fa-ban", "hidden": true, - "default": "False", + "default": false, "help_text": "Do not load `igenomes.config` when running the pipeline. You may choose this option if you observe clashes between custom parameters and those supplied in `igenomes.config`." } }, @@ -128,44 +114,42 @@ }, "Adapter trimming options": { "type": "object", - "description": "", - "default": "", "properties": { "clip_r1": { "type": "integer", - "default": "0", + "default": 0, "description": "Instructs Trim Galore to remove bp from the 5' end of read 1 (or single-end reads)" }, "clip_r2": { "type": "integer", - "default": "0", + "default": 0, "description": "Instructs Trim Galore to remove bp from the 5' end of read 2 (paired-end reads only)" }, "three_prime_clip_r1": { "type": "integer", - "default": "0", + "default": 0, "description": "Instructs Trim Galore to remove bp from the 3' end of read 1 AFTER adapter/quality trimming has been performed" }, "three_prime_clip_r2": { "type": "integer", - "default": "0", + "default": 0, "description": "Instructs Trim Galore to remove bp from the 3' end of read 2 AFTER adapter/quality trimming has been performed" }, "trim_nextseq": { "type": "integer", - "default": "0", + "default": 0, "description": "Instructs Trim Galore to apply the --nextseq=X option, to trim based on quality after removing poly-G tails", "help_text": "This enables the option Cutadapt `--nextseq-trim=3'CUTOFF` option via Trim Galore, which will set a quality cutoff (that is normally given with -q instead), but qualities of G bases are ignored. This trimming is in common for the NextSeq- and NovaSeq-platforms, where basecalls without any signal are called as high-quality G bases." }, "skip_trimming": { "type": "boolean", - "default": "False", + "default": false, "description": "Skip the adapter trimming step", "help_text": "Skip the adapter trimming step. Use this if your input FastQ files have already been trimmed outside of the workflow or if you're very confident that there is no adapter contamination in your data." }, "save_trimmed": { "type": "boolean", - "default": "False", + "default": false, "description": "Save the trimmed FastQ files in the results directory", "help_text": "By default, trimmed FastQ files will not be saved to the results directory. Specify this flag (or set to true in your config file) to copy these files to the results directory when complete." } @@ -173,33 +157,31 @@ }, "Alignment options": { "type": "object", - "description": "", - "default": "", "properties": { "keep_mito": { "type": "boolean", - "default": "False", + "default": false, "description": "Reads mapping to mitochondrial contig are not filtered from alignments" }, "keep_dups": { "type": "boolean", - "default": "False", + "default": false, "description": "Duplicate reads are not filtered from alignments" }, "keep_multi_map": { "type": "boolean", - "default": "False", + "default": false, "description": "Reads mapping to multiple locations are not filtered from alignments" }, "skip_merge_replicates": { "type": "boolean", - "default": "False", + "default": false, "description": "Do not perform alignment merging and downstream analysis by merging replicates i.e. only do this by merging resequenced libraries", "help_text": "An additional series of steps are performed by the pipeline by merging the replicates from the same experimental group. This is primarily to increase the sequencing depth in order to perform downstream analyses such as footprinting. Specifying this parameter means that these steps will not be performed." }, "save_align_intermeds": { "type": "boolean", - "default": "False", + "default": false, "description": "Save the intermediate BAM files from the alignment step", "help_text": "By default, intermediate BAM files will not be saved. The final BAM files created after the appropriate filtering step are always saved to limit storage usage. Set this parameter to also save other intermediate BAM files." } @@ -207,114 +189,105 @@ }, "Peak calling options": { "type": "object", - "description": "", - "default": "", "properties": { "narrow_peak": { "type": "boolean", - "default": "False", + "default": false, "description": "Run MACS2 in narrowPeak mode", "help_text": "MACS2 is run by default with the [`--broad`](https://github.com/taoliu/MACS#--broad) flag. Specify this flag to call peaks in narrowPeak mode." }, "broad_cutoff": { "type": "number", - "default": "0.1", + "default": 0.1, "description": "Specifies broad cutoff value for MACS2. Only used when --narrow_peak isnt specified", "help_text": "Specifies broad cut-off value for MACS2. Only used when `--narrow_peak` isnt specified." }, "min_reps_consensus": { "type": "integer", - "default": "1", + "default": 1, "description": "Number of biological replicates required from a given condition for a peak to contribute to a consensus peak", "help_text": "Number of biological replicates required from a given condition for a peak to contribute to a consensus peak . If you are confident you have good reproducibility amongst your replicates then you can increase the value of this parameter to create a \"reproducible\" set of consensus of peaks. For example, a value of 2 will mean peaks that have been called in at least 2 replicates will contribute to the consensus set of peaks, and as such peaks that are unique to a given replicate will be discarded." }, "save_macs_pileup": { "type": "boolean", - "default": "False", + "default": false, "description": "Instruct MACS2 to create bedGraph files normalised to signal per million reads" }, "skip_consensus_peaks": { "type": "boolean", - "default": "False", + "default": false, "description": "Skip consensus peak generation, annotation and counting" } } }, "Differential analysis options": { "type": "object", - "description": "", - "default": "", "properties": { "deseq2_vst": { "type": "boolean", - "default": "False", + "default": false, "description": "Use vst transformation instead of rlog with DESeq2", "help_text": "Use `vst` transformation instead of `rlog` with DESeq2. See [DESeq2 docs](http://bioconductor.org/packages/devel/bioc/vignettes/DESeq2/inst/doc/DESeq2.html#data-transformations-and-visualization)." }, "skip_diff_analysis": { "type": "boolean", - "default": "False", + "default": false, "description": "Skip differential accessibility analysis" } } }, "Process skipping options": { "type": "object", - "description": "", - "default": "", "properties": { "skip_fastqc": { "type": "boolean", - "default": "False", + "default": false, "description": "Skip FastQC" }, "skip_picard_metrics": { "type": "boolean", - "default": "False", + "default": false, "description": "Skip Picard CollectMultipleMetrics" }, "skip_preseq": { "type": "boolean", - "default": "False", + "default": false, "description": "Skip Preseq" }, "skip_plot_profile": { "type": "boolean", - "default": "False", + "default": false, "description": "Skip deepTools plotProfile" }, "skip_plot_fingerprint": { "type": "boolean", - "default": "False", + "default": false, "description": "Skip deepTools plotFingerprint" }, "skip_ataqv": { "type": "boolean", - "default": "False", + "default": false, "description": "Skip Ataqv" }, "skip_igv": { "type": "boolean", - "default": "False", + "default": false, "description": "Skip IGV" }, "skip_multiqc": { "type": "boolean", - "default": "False", + "default": false, "description": "Skip MultiQC" } } }, "Pipeline config options": { "type": "object", - "description": "", - "default": "", "properties": { "multiqc_config": { "type": "string", "description": "Custom config file to supply to MultiQC", - "default": "false", - "fa_icon": "", + "fa_icon": "fas fa-cog", "hidden": true }, "bamtools_filter_pe_config": { @@ -333,8 +306,6 @@ }, "Institutional config options": { "type": "object", - "description": "", - "default": "", "properties": { "custom_config_version": { "type": "string", @@ -352,39 +323,33 @@ "hostnames": { "type": "string", "description": "Institutional configs hostname", - "default": "false", "hidden": true }, "config_profile_description": { "type": "string", "description": "Institutional config description", "hidden": true, - "default": "false" }, "config_profile_contact": { "type": "string", "description": "Institutional config contact information", "hidden": true, - "default": "false" }, "config_profile_url": { "type": "string", "description": "Institutional config URL link", "hidden": true, - "default": "false" } } }, "Max job request options": { "type": "object", - "description": "", - "default": "", "properties": { "max_cpus": { "type": "integer", "description": "Maximum number of CPUs that can be requested for any single job", "default": 16, - "fa_icon": "", + "fa_icon": "fas fa-microchip", "hidden": true, "help_text": "Use to set an upper-limit for the CPU requirement for each process. Should be an integer e.g. `--max_cpus 1`" }, @@ -392,7 +357,7 @@ "type": "string", "description": "Maximum amount of memory that can be requested for any single job", "default": "128 GB", - "fa_icon": "", + "fa_icon": "fas fa-memory", "hidden": true, "help_text": "Use to set an upper-limit for the memory requirement for each process. Should be a string in the format integer-unit e.g. `--max_memory '8.GB'`" }, @@ -400,7 +365,7 @@ "type": "string", "description": "Maximum amount of time that can be requested for any single job", "default": "10d", - "fa_icon": "", + "fa_icon": "far fa-clock", "hidden": true, "help_text": "Use to set an upper-limit for the time requirement for each process. Should be a string in the format integer-unit e.g. `--max_time '2.h'`" } @@ -408,21 +373,19 @@ }, "Pipeline template options": { "type": "object", - "description": "", - "default": "", "properties": { "help": { "type": "boolean", "description": "Display help text", "hidden": true, - "fa_icon": "", - "default": "False" + "fa_icon": "fas fa-question-circle", + "default": false }, "outdir": { "type": "string", "description": "The output directory where the results will be saved", "default": "./results", - "fa_icon": "" + "fa_icon": "fas fa-folder-open" }, "publish_dir_mode": { "type": "string", @@ -430,7 +393,7 @@ "hidden": true, "description": "Method used to save pipeline results to output directory", "help_text": "The Nextflow `publishDir` option specifies which intermediate files should be saved to the output directory. This option tells the pipeline what method should be used to move these files. See https://www.nextflow.io/docs/latest/process.html#publishdir for details.", - "fa_icon": "", + "fa_icon": "fas fa-copy", "enum": [ "symlink", "rellink", @@ -443,55 +406,53 @@ "name": { "type": "string", "description": "Workflow name", - "fa_icon": "", + "fa_icon": "fas fa-fingerprint", "help_text": "A custom name for the pipeline run. Unlike the core nextflow `-name` option with one hyphen this parameter can be reused multiple times, for example if using `-resume`. Passed through to steps such as MultiQC and used for things like report filenames and titles.", - "default": "false" }, "email": { "type": "string", "description": "Email address for completion summary", - "fa_icon": "", + "fa_icon": "fas fa-envelope", "help_text": "An email address to send a summary email to when the pipeline is completed.", "pattern": "^([a-zA-Z0-9_\\-\\.]+)@([a-zA-Z0-9_\\-\\.]+)\\.([a-zA-Z]{2,5})$" }, "email_on_fail": { "type": "string", "description": "Email address for completion summary, only when pipeline fails", - "fa_icon": "", + "fa_icon": "fas fa-exclamation-triangle", "pattern": "^([a-zA-Z0-9_\\-\\.]+)@([a-zA-Z0-9_\\-\\.]+)\\.([a-zA-Z]{2,5})$", "help_text": "An email address to send a summary email to when the pipeline is completed - ONLY sent if the pipeline does not exit successfully." }, "plaintext_email": { "type": "boolean", "description": "Send plain-text email instead of HTML", - "fa_icon": "", + "fa_icon": "fas fa-remove-format", "hidden": true, - "default": "False" + "default": false }, "max_multiqc_email_size": { "type": "string", "description": "File size limit when attaching MultiQC reports to summary emails", "default": "25 MB", - "fa_icon": "", + "fa_icon": "fas fa-file-upload", "hidden": true }, "monochrome_logs": { "type": "boolean", "description": "Do not use coloured log outputs", - "fa_icon": "", + "fa_icon": "fas fa-palette", "hidden": true, - "default": "False" + "default": false }, "tracedir": { "type": "string", "description": "Directory to keep pipeline Nextflow logs and reports", "default": "\"${params.outdir}/pipeline_info\"", - "fa_icon": "", + "fa_icon": "fas fa-cogs", "hidden": true }, "clusterOptions": { "type": "string", - "default": "false", "description": "Arguments passed to Nextflow clusterOptions", "hidden": true } From cb7a0a2df4f1e85a062e43392d67becad4656867 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 25 Mar 2020 14:43:18 +0000 Subject: [PATCH 146/226] Fix schema with latest changes --- nextflow_schema.json | 312 +++++++++++++++++++++++++++---------------- 1 file changed, 198 insertions(+), 114 deletions(-) diff --git a/nextflow_schema.json b/nextflow_schema.json index 767d5691..2cf59615 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -11,96 +11,116 @@ "input": { "type": "string", "default": "./design.csv", - "description": "Comma-separated file containing information about the samples in the experiment", - "help_text": "You will need to create a design file with information about the samples in your experiment before running the pipeline. Use this parameter to specify its location. It has to be a comma-separated file with 4 columns, and a header row. See [usage docs](https://nf-co.re/atacseq/docs/usage#--input)." + "description": "Comma-separated file containing information about the samples in the experiment.", + "help_text": "You will need to create a design file with information about the samples in your experiment before running the pipeline. Use this parameter to specify its location. It has to be a comma-separated file with 4 columns, and a header row. See [usage docs](https://nf-co.re/atacseq/docs/usage#--input).", + "fa_icon": "fas fa-file-csv" }, "single_end": { "type": "boolean", - "description": "Specifies that the input is single-end reads", + "description": "Specifies that the input is single-end reads.", "fa_icon": "fas fa-align-center", "default": false, - "help_text": "By default, the pipeline expects paired-end data. If you have single-end data, specify `--single_end` on the command line when you launch the pipeline. It is not possible to run a mixture of single-end and paired-end files in one run." + "help_text": "By default, the pipeline expects paired-end data. If you have single-end data, specify this parameter on the command line when you launch the pipeline. It is not possible to run a mixture of single-end and paired-end files in one run." }, "fragment_size": { "type": "integer", "default": 0, - "description": "Estimated fragment size used to extend single-end reads" + "description": "Estimated fragment size used to extend single-end reads.", + "fa_icon": "fas fa-align-justify", + "help_text": "" }, "fingerprint_bins": { "type": "integer", "default": 500000, - "description": "Number of genomic bins to use when calculating deepTools fingerprint plot" + "description": "Number of genomic bins to use when calculating deepTools fingerprint plot.", + "fa_icon": "fas fa-dumpster" }, "seq_center": { "type": "string", - "description": "Sequencing center information to be added to read group of BAM files" + "description": "Sequencing center information to be added to read group of BAM files.", + "fa_icon": "fas fa-synagogue" } }, "required": [ "input" - ] + ], + "fa_icon": "fas fa-terminal" }, "Reference genome options": { "type": "object", "properties": { "genome": { "type": "string", - "description": "Name of iGenomes reference", + "description": "Name of iGenomes reference.", "fa_icon": "fas fa-book", - "help_text": "If using a reference genome configured in the pipeline using iGenomes, use this parameter to give the ID for the reference. This is then used to build the full paths for all required reference genome files. For example: `--genome GRCh38`.", + "help_text": "If using a reference genome configured in the pipeline using iGenomes, use this parameter to give the ID for the reference. This is then used to build the full paths for all required reference genome files e.g. `--genome GRCh38`." }, "fasta": { "type": "string", - "description": "Path to Fasta reference. Not mandatory when using reference in iGenomes config via --genome", - "help_text": "Full path to fasta file containing reference genome (*mandatory* if `--genome` is not specified). If you don't have a BWA index available this will be generated for you automatically. Combine with `--save_reference` to save BWA index for future runs." + "description": "Path to Fasta reference file.", + "help_text": "This parameter is *mandatory* if `--genome` is not specified. If you don't have a BWA index available this will be generated for you automatically. Combine with `--save_reference` to save BWA index for future runs.", + "fa_icon": "far fa-file-code" }, "gtf": { "type": "string", - "description": "Path to GTF file. Not mandatory when using reference in iGenomes config via --genome" + "description": "Path to GTF annotation file.", + "fa_icon": "fas fa-file-invoice", + "help_text": "This parameter is *mandatory* if `--genome` is not specified." }, "bwa_index": { "type": "string", - "description": "Full path to directory containing BWA index including base name i.e. /path/to/index/genome.fa" + "description": "Full path to directory containing BWA index including base name i.e. /path/to/index/genome.fa.", + "fa_icon": "fas fa-bezier-curve", + "help_text": "" }, "gene_bed": { "type": "string", - "description": "Path to BED file containing gene intervals. This will be created from the GTF file if not specified." + "description": "Path to BED file containing gene intervals. This will be created from the GTF file if not specified.", + "fa_icon": "fas fa-procedures", + "help_text": "" }, "tss_bed": { "type": "string", - "description": "Path to BED file containing transcription start sites. This will be created from the gene BED file if not specified." + "description": "Path to BED file containing transcription start sites. This will be created from the gene BED file if not specified.", + "fa_icon": "fas fa-procedures", + "help_text": "" }, "macs_gsize": { "type": "string", - "description": "Effective genome size parameter required by MACS2. If using iGenomes config, values have only been provided when --genome is set as GRCh37, GRCm38, hg19, mm10, BDGP6 and WBcel235", - "help_text": "[Effective genome size](https://github.com/taoliu/MACS#-g--gsize) parameter required by MACS2. These have been provided when `--genome` is set as *GRCh37*, *GRCh38*, *GRCm38*, *WBcel235*, *BDGP6*, *R64-1-1*, *EF2*, *hg38*, *hg19* and *mm10*. For other genomes, if this parameter is not specified then the MACS2 peak-calling and differential analysis will be skipped." + "description": "Effective genome size parameter required by MACS2.", + "help_text": "[Effective genome size](https://github.com/taoliu/MACS#-g--gsize) parameter required by MACS2. If using an iGenomes reference these have been provided when `--genome` is set as *GRCh37*, *GRCh38*, *GRCm38*, *WBcel235*, *BDGP6*, *R64-1-1*, *EF2*, *hg38*, *hg19* and *mm10*. For other genomes, if this parameter is not specified then the MACS2 peak-calling and differential analysis will be skipped.", + "fa_icon": "fas fa-arrows-alt-h" }, "blacklist": { "type": "string", - "description": "Path to blacklist regions (.BED format), used for filtering alignments", - "help_text": "If provided, alignments that overlap with the regions in this file will be filtered out (see [ENCODE blacklists](https://sites.google.com/site/anshulkundaje/projects/blacklists)). The file should be in BED format. Blacklisted regions for *GRCh37*, *GRCh38*, *GRCm38*, *hg19*, *hg38*, *mm10* are bundled with the pipeline in the [`blacklists`](../assets/blacklists/) directory, and as such will be automatically used if any of those genomes are specified with the `--genome` parameter." + "description": "Path to blacklist regions in BED format, used for filtering alignments.", + "help_text": "If provided, alignments that overlap with the regions in this file will be filtered out (see [ENCODE blacklists](https://sites.google.com/site/anshulkundaje/projects/blacklists)). The file should be in BED format. Blacklisted regions for *GRCh37*, *GRCh38*, *GRCm38*, *hg19*, *hg38*, *mm10* are bundled with the pipeline in the [`blacklists`](../assets/blacklists/) directory, and as such will be automatically used if any of those genomes are specified with the `--genome` parameter.", + "fa_icon": "fas fa-clipboard-list" }, "mito_name": { "type": "string", - "description": "Name of Mitochondrial chomosome in genome fasta (e.g. chrM). Reads aligning to this contig are filtered out", - "help_text": "Name of mitochondrial chomosome in reference assembly. Reads aligning to this contig are filtered out if a valid identifier is provided otherwise this step is skipped. Where possible these have been provided in the [`igenomes.config`](../conf/igenomes.config)." + "description": "Name of Mitochondrial chomosome in reference assembly e.g. chrM.", + "help_text": "Reads aligning to this contig are filtered out if a valid identifier is provided otherwise this step is skipped. Where possible these have been provided in the [`igenomes.config`](../conf/igenomes.config).", + "fa_icon": "fas fa-signature" }, "save_reference": { "type": "boolean", "default": false, - "description": "If generated by the pipeline save the BWA index in the results directory", - "help_text": "If the BWA index is generated by the pipeline use this parameter to save it to your results folder. These can then be used for future pipeline runs, reducing processing times." + "description": "If generated by the pipeline save the BWA index in the results directory.", + "help_text": "If the BWA index is generated by the pipeline use this parameter to save it to your results folder. These can then be used for future pipeline runs, reducing processing times.", + "fa_icon": "fas fa-save" }, "igenomes_base": { "type": "string", - "description": "Directory / URL base for iGenomes references", + "description": "Directory / URL base for iGenomes references.", "default": "s3://ngi-igenomes/igenomes/", "fa_icon": "fas fa-cloud-download-alt", - "hidden": true + "hidden": true, + "help_text": "" }, "igenomes_ignore": { "type": "boolean", - "description": "Do not load the iGenomes reference config", + "description": "Do not load the iGenomes reference config.", "fa_icon": "fas fa-ban", "hidden": true, "default": false, @@ -110,7 +130,8 @@ "required": [ "fasta", "gtf" - ] + ], + "fa_icon": "fas fa-dna" }, "Adapter trimming options": { "type": "object", @@ -118,42 +139,52 @@ "clip_r1": { "type": "integer", "default": 0, - "description": "Instructs Trim Galore to remove bp from the 5' end of read 1 (or single-end reads)" + "description": "Instructs Trim Galore to remove bp from the 5' end of read 1 (or single-end reads).", + "fa_icon": "fas fa-cut", + "help_text": "" }, "clip_r2": { "type": "integer", "default": 0, - "description": "Instructs Trim Galore to remove bp from the 5' end of read 2 (paired-end reads only)" + "description": "Instructs Trim Galore to remove bp from the 5' end of read 2 (paired-end reads only).", + "fa_icon": "fas fa-cut", + "help_text": "" }, "three_prime_clip_r1": { "type": "integer", "default": 0, - "description": "Instructs Trim Galore to remove bp from the 3' end of read 1 AFTER adapter/quality trimming has been performed" + "description": "Instructs Trim Galore to remove bp from the 3' end of read 1 AFTER adapter/quality trimming has been performed.", + "fa_icon": "fas fa-cut" }, "three_prime_clip_r2": { "type": "integer", "default": 0, - "description": "Instructs Trim Galore to remove bp from the 3' end of read 2 AFTER adapter/quality trimming has been performed" + "description": "Instructs Trim Galore to remove bp from the 3' end of read 2 AFTER adapter/quality trimming has been performed.", + "fa_icon": "fas fa-cut" }, "trim_nextseq": { "type": "integer", "default": 0, - "description": "Instructs Trim Galore to apply the --nextseq=X option, to trim based on quality after removing poly-G tails", - "help_text": "This enables the option Cutadapt `--nextseq-trim=3'CUTOFF` option via Trim Galore, which will set a quality cutoff (that is normally given with -q instead), but qualities of G bases are ignored. This trimming is in common for the NextSeq- and NovaSeq-platforms, where basecalls without any signal are called as high-quality G bases." + "description": "Instructs Trim Galore to apply the --nextseq=X option, to trim based on quality after removing poly-G tails.", + "help_text": "This enables the option Cutadapt `--nextseq-trim=3'CUTOFF` option via Trim Galore, which will set a quality cutoff (that is normally given with -q instead), but qualities of G bases are ignored. This trimming is in common for the NextSeq- and NovaSeq-platforms, where basecalls without any signal are called as high-quality G bases.", + "fa_icon": "fas fa-cut" }, "skip_trimming": { "type": "boolean", "default": false, - "description": "Skip the adapter trimming step", - "help_text": "Skip the adapter trimming step. Use this if your input FastQ files have already been trimmed outside of the workflow or if you're very confident that there is no adapter contamination in your data." + "description": "Skip the adapter trimming step.", + "help_text": "Use this if your input FastQ files have already been trimmed outside of the workflow or if you're very confident that there is no adapter contamination in your data.", + "fa_icon": "fas fa-forward" }, "save_trimmed": { "type": "boolean", "default": false, - "description": "Save the trimmed FastQ files in the results directory", - "help_text": "By default, trimmed FastQ files will not be saved to the results directory. Specify this flag (or set to true in your config file) to copy these files to the results directory when complete." + "description": "Save the trimmed FastQ files in the results directory.", + "help_text": "By default, trimmed FastQ files will not be saved to the results directory. Specify this flag (or set to true in your config file) to copy these files to the results directory when complete.", + "fa_icon": "fas fa-save" } - } + }, + "fa_icon": "fas fa-cut" }, "Alignment options": { "type": "object", @@ -161,31 +192,37 @@ "keep_mito": { "type": "boolean", "default": false, - "description": "Reads mapping to mitochondrial contig are not filtered from alignments" + "description": "Reads mapping to mitochondrial contig are not filtered from alignments.", + "fa_icon": "fas fa-cart-arrow-down" }, "keep_dups": { "type": "boolean", "default": false, - "description": "Duplicate reads are not filtered from alignments" + "description": "Duplicate reads are not filtered from alignments.", + "fa_icon": "fas fa-cart-arrow-down" }, "keep_multi_map": { "type": "boolean", "default": false, - "description": "Reads mapping to multiple locations are not filtered from alignments" + "description": "Reads mapping to multiple locations are not filtered from alignments.", + "fa_icon": "fas fa-cart-arrow-down" }, "skip_merge_replicates": { "type": "boolean", "default": false, - "description": "Do not perform alignment merging and downstream analysis by merging replicates i.e. only do this by merging resequenced libraries", - "help_text": "An additional series of steps are performed by the pipeline by merging the replicates from the same experimental group. This is primarily to increase the sequencing depth in order to perform downstream analyses such as footprinting. Specifying this parameter means that these steps will not be performed." + "description": "Do not perform alignment merging and downstream analysis by merging replicates i.e. only do this by merging resequenced libraries.", + "help_text": "An additional series of steps are performed by the pipeline for merging the replicates from the same experimental group. This is primarily to increase the sequencing depth in order to perform downstream analyses such as footprinting. Specifying this parameter means that these steps will not be performed.", + "fa_icon": "fas fa-forward" }, "save_align_intermeds": { "type": "boolean", "default": false, - "description": "Save the intermediate BAM files from the alignment step", - "help_text": "By default, intermediate BAM files will not be saved. The final BAM files created after the appropriate filtering step are always saved to limit storage usage. Set this parameter to also save other intermediate BAM files." + "description": "Save the intermediate BAM files from the alignment step.", + "help_text": "By default, intermediate BAM files will not be saved. The final BAM files created after the appropriate filtering step are always saved to limit storage usage. Set this parameter to also save other intermediate BAM files.", + "fa_icon": "fas fa-save" } - } + }, + "fa_icon": "fas fa-map-signs" }, "Peak calling options": { "type": "object", @@ -193,32 +230,39 @@ "narrow_peak": { "type": "boolean", "default": false, - "description": "Run MACS2 in narrowPeak mode", - "help_text": "MACS2 is run by default with the [`--broad`](https://github.com/taoliu/MACS#--broad) flag. Specify this flag to call peaks in narrowPeak mode." + "description": "Run MACS2 in narrowPeak mode.", + "help_text": "MACS2 is run by default with the [`--broad`](https://github.com/taoliu/MACS#--broad) flag. Specify this flag to call peaks in narrowPeak mode.", + "fa_icon": "fas fa-arrows-alt-h" }, "broad_cutoff": { "type": "number", "default": 0.1, - "description": "Specifies broad cutoff value for MACS2. Only used when --narrow_peak isnt specified", - "help_text": "Specifies broad cut-off value for MACS2. Only used when `--narrow_peak` isnt specified." + "description": "Specifies broad cutoff value for MACS2. Only used when --narrow_peak isnt specified.", + "fa_icon": "fas fa-hand-scissors" }, "min_reps_consensus": { "type": "integer", "default": 1, - "description": "Number of biological replicates required from a given condition for a peak to contribute to a consensus peak", - "help_text": "Number of biological replicates required from a given condition for a peak to contribute to a consensus peak . If you are confident you have good reproducibility amongst your replicates then you can increase the value of this parameter to create a \"reproducible\" set of consensus of peaks. For example, a value of 2 will mean peaks that have been called in at least 2 replicates will contribute to the consensus set of peaks, and as such peaks that are unique to a given replicate will be discarded." + "description": "Number of biological replicates required from a given condition for a peak to contribute to a consensus peak.", + "help_text": "If you are confident you have good reproducibility amongst your replicates then you can increase the value of this parameter to create a \"reproducible\" set of consensus peaks. For example, a value of 2 will mean peaks that have been called in at least 2 replicates will contribute to the consensus set of peaks, and as such peaks that are unique to a given replicate will be discarded.", + "fa_icon": "fas fa-sort-numeric-down" }, "save_macs_pileup": { "type": "boolean", "default": false, - "description": "Instruct MACS2 to create bedGraph files normalised to signal per million reads" + "description": "Instruct MACS2 to create bedGraph files normalised to signal per million reads.", + "fa_icon": "fas fa-save", + "help_text": "" }, "skip_consensus_peaks": { "type": "boolean", "default": false, - "description": "Skip consensus peak generation, annotation and counting" + "description": "Skip consensus peak generation, annotation and counting.", + "fa_icon": "fas fa-forward", + "help_text": "" } - } + }, + "fa_icon": "fas fa-chart-area" }, "Differential analysis options": { "type": "object", @@ -226,15 +270,19 @@ "deseq2_vst": { "type": "boolean", "default": false, - "description": "Use vst transformation instead of rlog with DESeq2", - "help_text": "Use `vst` transformation instead of `rlog` with DESeq2. See [DESeq2 docs](http://bioconductor.org/packages/devel/bioc/vignettes/DESeq2/inst/doc/DESeq2.html#data-transformations-and-visualization)." + "description": "Use vst transformation instead of rlog with DESeq2.", + "help_text": "See [DESeq2 docs](http://bioconductor.org/packages/devel/bioc/vignettes/DESeq2/inst/doc/DESeq2.html#data-transformations-and-visualization).", + "fa_icon": "fas fa-dolly" }, "skip_diff_analysis": { "type": "boolean", "default": false, - "description": "Skip differential accessibility analysis" + "description": "Skip differential accessibility analysis.", + "fa_icon": "fas fa-forward", + "help_text": "" } - } + }, + "fa_icon": "fas fa-not-equal" }, "Process skipping options": { "type": "object", @@ -242,112 +290,139 @@ "skip_fastqc": { "type": "boolean", "default": false, - "description": "Skip FastQC" + "description": "Skip FastQC.", + "fa_icon": "fas fa-forward" }, "skip_picard_metrics": { "type": "boolean", "default": false, - "description": "Skip Picard CollectMultipleMetrics" + "description": "Skip Picard CollectMultipleMetrics.", + "fa_icon": "fas fa-forward" }, "skip_preseq": { "type": "boolean", "default": false, - "description": "Skip Preseq" + "description": "Skip Preseq.", + "fa_icon": "fas fa-forward" }, "skip_plot_profile": { "type": "boolean", "default": false, - "description": "Skip deepTools plotProfile" + "description": "Skip deepTools plotProfile.", + "fa_icon": "fas fa-forward" }, "skip_plot_fingerprint": { "type": "boolean", "default": false, - "description": "Skip deepTools plotFingerprint" + "description": "Skip deepTools plotFingerprint.", + "fa_icon": "fas fa-forward" }, "skip_ataqv": { "type": "boolean", "default": false, - "description": "Skip Ataqv" + "description": "Skip Ataqv.", + "fa_icon": "fas fa-forward" }, "skip_igv": { "type": "boolean", "default": false, - "description": "Skip IGV" + "description": "Skip IGV.", + "fa_icon": "fas fa-forward" }, "skip_multiqc": { "type": "boolean", "default": false, - "description": "Skip MultiQC" + "description": "Skip MultiQC.", + "fa_icon": "fas fa-forward" } - } + }, + "fa_icon": "fas fa-forward" }, "Pipeline config options": { "type": "object", "properties": { "multiqc_config": { "type": "string", - "description": "Custom config file to supply to MultiQC", + "description": "Custom config file to supply to MultiQC.", "fa_icon": "fas fa-cog", - "hidden": true + "hidden": true, + "help_text": "" }, "bamtools_filter_pe_config": { "type": "string", - "default": "\"$baseDir/assets/bamtools_filter_pe.json\"", + "default": "$baseDir/assets/bamtools_filter_pe.json", "hidden": true, - "description": "BAMTools JSON file with custom filters for paired-end data" + "description": "BAMTools JSON file with custom filters for paired-end data.", + "fa_icon": "fas fa-cog", + "help_text": "" }, "bamtools_filter_se_config": { "type": "string", - "default": "\"$baseDir/assets/bamtools_filter_se.json\"", + "default": "$baseDir/assets/bamtools_filter_se.json", "hidden": true, - "description": "BAMTools JSON file with custom filters for single-end data" + "description": "BAMTools JSON file with custom filters for single-end data.", + "fa_icon": "fas fa-cog", + "help_text": "" } - } + }, + "fa_icon": "fas fa-cog" }, "Institutional config options": { "type": "object", "properties": { "custom_config_version": { "type": "string", - "description": "Git commit id for Institutional configs", + "description": "Git commit id for Institutional configs.", "default": "master", - "hidden": true + "hidden": true, + "fa_icon": "fas fa-users-cog", + "help_text": "" }, "custom_config_base": { "type": "string", - "description": "Base directory for Institutional configs", + "description": "Base directory for Institutional configs.", "default": "https://raw.githubusercontent.com/nf-core/configs/master", "hidden": true, - "help_text": "If you're running offline, Nextflow will not be able to fetch the institutional config files\nfrom the internet. If you don't need them, then this is not a problem. If you do need them,\nyou should download the files from the repo and tell nextflow where to find them with the `--custom_config_base` option." + "help_text": "If you're running offline, Nextflow will not be able to fetch the institutional config files from the internet. If you don't need them, then this is not a problem. If you do need them, you should download the files from the repo and tell Nextflow where to find them with this parameter.", + "fa_icon": "fas fa-users-cog" }, "hostnames": { "type": "string", - "description": "Institutional configs hostname", - "hidden": true + "description": "Institutional configs hostname.", + "hidden": true, + "fa_icon": "fas fa-users-cog", + "help_text": "" }, "config_profile_description": { "type": "string", - "description": "Institutional config description", + "description": "Institutional config description.", "hidden": true, + "fa_icon": "fas fa-users-cog", + "help_text": "" }, "config_profile_contact": { "type": "string", - "description": "Institutional config contact information", + "description": "Institutional config contact information.", "hidden": true, + "fa_icon": "fas fa-users-cog", + "help_text": "" }, "config_profile_url": { "type": "string", - "description": "Institutional config URL link", + "description": "Institutional config URL link.", "hidden": true, + "fa_icon": "fas fa-users-cog", + "help_text": "" } - } + }, + "fa_icon": "fas fa-university" }, "Max job request options": { "type": "object", "properties": { "max_cpus": { "type": "integer", - "description": "Maximum number of CPUs that can be requested for any single job", + "description": "Maximum number of CPUs that can be requested for any single job.", "default": 16, "fa_icon": "fas fa-microchip", "hidden": true, @@ -355,44 +430,46 @@ }, "max_memory": { "type": "string", - "description": "Maximum amount of memory that can be requested for any single job", - "default": "128 GB", + "description": "Maximum amount of memory that can be requested for any single job.", + "default": "128.GB", "fa_icon": "fas fa-memory", "hidden": true, "help_text": "Use to set an upper-limit for the memory requirement for each process. Should be a string in the format integer-unit e.g. `--max_memory '8.GB'`" }, "max_time": { "type": "string", - "description": "Maximum amount of time that can be requested for any single job", - "default": "10d", + "description": "Maximum amount of time that can be requested for any single job.", + "default": "240.h", "fa_icon": "far fa-clock", "hidden": true, "help_text": "Use to set an upper-limit for the time requirement for each process. Should be a string in the format integer-unit e.g. `--max_time '2.h'`" } - } + }, + "fa_icon": "fab fa-acquisitions-incorporated" }, "Pipeline template options": { "type": "object", "properties": { "help": { "type": "boolean", - "description": "Display help text", + "description": "Display help text.", "hidden": true, "fa_icon": "fas fa-question-circle", "default": false }, "outdir": { "type": "string", - "description": "The output directory where the results will be saved", + "description": "The output directory where the results will be saved.", "default": "./results", - "fa_icon": "fas fa-folder-open" + "fa_icon": "fas fa-folder-open", + "help_text": "" }, "publish_dir_mode": { "type": "string", "default": "copy", "hidden": true, - "description": "Method used to save pipeline results to output directory", - "help_text": "The Nextflow `publishDir` option specifies which intermediate files should be saved to the output directory. This option tells the pipeline what method should be used to move these files. See https://www.nextflow.io/docs/latest/process.html#publishdir for details.", + "description": "Method used to save pipeline results to output directory.", + "help_text": "The Nextflow `publishDir` option specifies which intermediate files should be saved to the output directory. This option tells the pipeline what method should be used to move these files. See [Nextflow docs](https://www.nextflow.io/docs/latest/process.html#publishdir) for details.", "fa_icon": "fas fa-copy", "enum": [ "symlink", @@ -405,58 +482,65 @@ }, "name": { "type": "string", - "description": "Workflow name", + "description": "Workflow name.", "fa_icon": "fas fa-fingerprint", - "help_text": "A custom name for the pipeline run. Unlike the core nextflow `-name` option with one hyphen this parameter can be reused multiple times, for example if using `-resume`. Passed through to steps such as MultiQC and used for things like report filenames and titles.", + "help_text": "A custom name for the pipeline run. Unlike the core nextflow `-name` option with one hyphen this parameter can be reused multiple times, for example if using `-resume`. Passed through to steps such as MultiQC and used for things like report filenames and titles." }, "email": { "type": "string", - "description": "Email address for completion summary", + "description": "Email address for completion summary.", "fa_icon": "fas fa-envelope", "help_text": "An email address to send a summary email to when the pipeline is completed.", "pattern": "^([a-zA-Z0-9_\\-\\.]+)@([a-zA-Z0-9_\\-\\.]+)\\.([a-zA-Z]{2,5})$" }, "email_on_fail": { "type": "string", - "description": "Email address for completion summary, only when pipeline fails", + "description": "Email address for completion summary, only when pipeline fails.", "fa_icon": "fas fa-exclamation-triangle", "pattern": "^([a-zA-Z0-9_\\-\\.]+)@([a-zA-Z0-9_\\-\\.]+)\\.([a-zA-Z]{2,5})$", "help_text": "An email address to send a summary email to when the pipeline is completed - ONLY sent if the pipeline does not exit successfully." }, "plaintext_email": { "type": "boolean", - "description": "Send plain-text email instead of HTML", + "description": "Send plain-text email instead of HTML.", "fa_icon": "fas fa-remove-format", "hidden": true, - "default": false + "default": false, + "help_text": "" }, "max_multiqc_email_size": { "type": "string", - "description": "File size limit when attaching MultiQC reports to summary emails", - "default": "25 MB", + "description": "File size limit when attaching MultiQC reports to summary emails.", + "default": "25.MB", "fa_icon": "fas fa-file-upload", - "hidden": true + "hidden": true, + "help_text": "" }, "monochrome_logs": { "type": "boolean", - "description": "Do not use coloured log outputs", + "description": "Do not use coloured log outputs.", "fa_icon": "fas fa-palette", "hidden": true, - "default": false + "default": false, + "help_text": "" }, "tracedir": { "type": "string", - "description": "Directory to keep pipeline Nextflow logs and reports", - "default": "\"${params.outdir}/pipeline_info\"", + "description": "Directory to keep pipeline Nextflow logs and reports.", + "default": "${params.outdir}/pipeline_info", "fa_icon": "fas fa-cogs", - "hidden": true + "hidden": true, + "help_text": "" }, "clusterOptions": { "type": "string", - "description": "Arguments passed to Nextflow clusterOptions", - "hidden": true + "description": "Arguments passed to Nextflow clusterOptions.", + "hidden": true, + "fa_icon": "fas fa-network-wired", + "help_text": "" } - } + }, + "fa_icon": "fas fa-file-import" } } } From 9fd388b68a901fd0a1a5df45eed4a21312095cc0 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 25 Mar 2020 16:34:24 +0000 Subject: [PATCH 147/226] Remove required field --- nextflow_schema.json | 4 ---- 1 file changed, 4 deletions(-) diff --git a/nextflow_schema.json b/nextflow_schema.json index 2cf59615..4487fd5e 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -127,10 +127,6 @@ "help_text": "Do not load `igenomes.config` when running the pipeline. You may choose this option if you observe clashes between custom parameters and those supplied in `igenomes.config`." } }, - "required": [ - "fasta", - "gtf" - ], "fa_icon": "fas fa-dna" }, "Adapter trimming options": { From 3d8e305d80f62a3c6a92854c5fe8eadbd66fb661 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 25 Mar 2020 17:54:36 +0000 Subject: [PATCH 148/226] Add PE param to preseq --- main.nf | 11 ++++++++++- 1 file changed, 10 insertions(+), 1 deletion(-) diff --git a/main.nf b/main.nf index 185138b4..563b2bc0 100644 --- a/main.nf +++ b/main.nf @@ -897,11 +897,20 @@ process MergedLibPreseq { output: file "*.ccurve.txt" into ch_mlib_preseq_mqc + file "*.log" into ch_mlib_preseq_log script: prefix = "${name}.mLb.mkD" + pe = params.single_end ? "" : "-pe" """ - preseq lc_extrap -v -output ${prefix}.ccurve.txt -bam ${bam[0]} + preseq lc_extrap \\ + -output ${prefix}.ccurve.txt \\ + -verbose \\ + -bam \\ + $pe \\ + -seed 1 \\ + ${bam[0]} + cp .command.err ${prefix}.command.log """ } From 7749ebc2e3a4123eea9d88d7b68013c1545d97d3 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 25 Mar 2020 18:02:51 +0000 Subject: [PATCH 149/226] Update CHANGELOG --- CHANGELOG.md | 1 + 1 file changed, 1 insertion(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index 07410391..d23a798a 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -13,6 +13,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * [#76](https://github.com/nf-core/atacseq/issues/76) - featureCounts coupled to DESeq2 * [#79](https://github.com/nf-core/atacseq/issues/79) - Parallelize DESeq2 * [#80](https://github.com/nf-core/atacseq/pull/80) - Added social preview image +* [#97](https://github.com/nf-core/atacseq/issues/97) - PBC1, PBC2 from pipeline? * Update template to tools `1.9` * Parameters: * `--skip_consensus_peaks` to skip consensus peak generation From 2947cc2de3f01e8b053b161df9631c55f5d1f69f Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 26 Mar 2020 12:13:32 +0000 Subject: [PATCH 150/226] Update nextflow.config --- nextflow.config | 0 1 file changed, 0 insertions(+), 0 deletions(-) mode change 100755 => 100644 nextflow.config diff --git a/nextflow.config b/nextflow.config old mode 100755 new mode 100644 From 999c8b3925de895a9951422730d18d677a29e4ac Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 26 Mar 2020 14:18:15 +0000 Subject: [PATCH 151/226] Add aws tests --- .github/workflows/awstest.yml | 31 +++++++++++++++++++++++++++++++ 1 file changed, 31 insertions(+) create mode 100644 .github/workflows/awstest.yml diff --git a/.github/workflows/awstest.yml b/.github/workflows/awstest.yml new file mode 100644 index 00000000..5bde6ff4 --- /dev/null +++ b/.github/workflows/awstest.yml @@ -0,0 +1,31 @@ +name: nf-core AWS test +# This workflow is triggered on PRs to the master branch. +# It runs the -profile 'test' on AWS batch + +on: + push: + branches: + - master + - dev # just for testing purposes, to be removed + release: + types: [published] + +jobs: + run-awstest: + name: Run AWS test + runs-on: ubuntu-latest + steps: + - name: Setup Miniconda + uses: goanpeca/setup-miniconda@v1.0.2 + with: + auto-update-conda: true + python-version: 3.7 + - name: Install awscli + run: conda install -c conda-forge awscli + - name: Start AWS batch job + env: + AWS_ACCESS_KEY_ID: ${{secrets.AWS_KEY_ID}} + AWS_SECRET_ACCESS_KEY: ${{secrets.AWS_KEY_SECRET}} + TOWER_ACCESS_TOKEN: ${{secrets.TOWER_ACCESS_TOKEN}} + run: | + aws batch submit-job --region eu-west-1 --job-name nf-core-atacseq --job-queue 'default-8b3836e0-5eda-11ea-96e5-0a2c3f6a2a32' --job-definition nextflow --container-overrides '{"command": ["nf-core/atacseq", "-r '"${GITHUB_SHA}"' -profile test --outdir s3://nf-core-awsmegatests/atacseq/results-'"${GITHUB_SHA}"' -w s3://nf-core-awsmegatests/atacseq/work-'"${GITHUB_SHA}"' -with-tower"], "environment": [{"name": "TOWER_ACCESS_TOKEN", "value": "'"$TOWER_ACCESS_TOKEN"'"}]}' From 21375791ccaf51cdadcbb03c14331b9873b03cfc Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 26 Mar 2020 17:54:12 +0000 Subject: [PATCH 152/226] Add test config for full test --- conf/test_full.config | 19 +++++++++++++++++++ 1 file changed, 19 insertions(+) create mode 100755 conf/test_full.config diff --git a/conf/test_full.config b/conf/test_full.config new file mode 100755 index 00000000..94cd8e5e --- /dev/null +++ b/conf/test_full.config @@ -0,0 +1,19 @@ +/* + * ------------------------------------------------- + * Nextflow config file for running tests + * ------------------------------------------------- + * Defines bundled input files and everything required + * to run a full pipeline test. Use as follows: + * nextflow run nf-core/atacseq -profile test_full, + */ + +params { + config_profile_name = 'Full test profile' + config_profile_description = 'Full test dataset to check pipeline function' + + // Input data + input = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/design_full.csv' + + // Genome references + genome = 'hg19' +} From 288030606ad83941937f5eb555d7dee6e2ec5e98 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 26 Mar 2020 17:54:22 +0000 Subject: [PATCH 153/226] Add profile for full test --- nextflow.config | 1 + 1 file changed, 1 insertion(+) diff --git a/nextflow.config b/nextflow.config index 9e16c428..2d5009de 100644 --- a/nextflow.config +++ b/nextflow.config @@ -122,6 +122,7 @@ profiles { singularity.autoMounts = true } test { includeConfig 'conf/test.config' } + test_full { includeConfig 'conf/test_full.config' } } // Load igenomes.config if required From 157c5189056b24a834c7a8b4bbc0bf8e52c82c19 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 26 Mar 2020 17:55:35 +0000 Subject: [PATCH 154/226] Change test to test_full --- .github/workflows/awstest.yml | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/.github/workflows/awstest.yml b/.github/workflows/awstest.yml index 5bde6ff4..ae7732b1 100644 --- a/.github/workflows/awstest.yml +++ b/.github/workflows/awstest.yml @@ -1,6 +1,6 @@ name: nf-core AWS test # This workflow is triggered on PRs to the master branch. -# It runs the -profile 'test' on AWS batch +# It runs the -profile 'test_full' on AWS batch on: push: @@ -28,4 +28,4 @@ jobs: AWS_SECRET_ACCESS_KEY: ${{secrets.AWS_KEY_SECRET}} TOWER_ACCESS_TOKEN: ${{secrets.TOWER_ACCESS_TOKEN}} run: | - aws batch submit-job --region eu-west-1 --job-name nf-core-atacseq --job-queue 'default-8b3836e0-5eda-11ea-96e5-0a2c3f6a2a32' --job-definition nextflow --container-overrides '{"command": ["nf-core/atacseq", "-r '"${GITHUB_SHA}"' -profile test --outdir s3://nf-core-awsmegatests/atacseq/results-'"${GITHUB_SHA}"' -w s3://nf-core-awsmegatests/atacseq/work-'"${GITHUB_SHA}"' -with-tower"], "environment": [{"name": "TOWER_ACCESS_TOKEN", "value": "'"$TOWER_ACCESS_TOKEN"'"}]}' + aws batch submit-job --region eu-west-1 --job-name nf-core-atacseq --job-queue 'default-8b3836e0-5eda-11ea-96e5-0a2c3f6a2a32' --job-definition nextflow --container-overrides '{"command": ["nf-core/atacseq", "-r '"${GITHUB_SHA}"' -profile test_full --outdir s3://nf-core-awsmegatests/atacseq/results-'"${GITHUB_SHA}"' -w s3://nf-core-awsmegatests/atacseq/work-'"${GITHUB_SHA}"' -with-tower"], "environment": [{"name": "TOWER_ACCESS_TOKEN", "value": "'"$TOWER_ACCESS_TOKEN"'"}]}' From 61d22ad1207f19cbe89b0cefb530e6e42e59340c Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 27 Mar 2020 11:01:58 +0000 Subject: [PATCH 155/226] Test on PR instead of push --- .github/workflows/awstest.yml | 5 +++-- 1 file changed, 3 insertions(+), 2 deletions(-) diff --git a/.github/workflows/awstest.yml b/.github/workflows/awstest.yml index ae7732b1..8902016a 100644 --- a/.github/workflows/awstest.yml +++ b/.github/workflows/awstest.yml @@ -3,10 +3,11 @@ name: nf-core AWS test # It runs the -profile 'test_full' on AWS batch on: - push: + #push: + pull_request: branches: - master - - dev # just for testing purposes, to be removed + #- dev # just for testing purposes, to be removed release: types: [published] From c0db0ff4d699b368789213aeb7d26d331b1e60ea Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 27 Mar 2020 11:07:55 +0000 Subject: [PATCH 156/226] Remove comments --- .github/workflows/awstest.yml | 2 -- 1 file changed, 2 deletions(-) diff --git a/.github/workflows/awstest.yml b/.github/workflows/awstest.yml index 8902016a..b7a47e78 100644 --- a/.github/workflows/awstest.yml +++ b/.github/workflows/awstest.yml @@ -3,11 +3,9 @@ name: nf-core AWS test # It runs the -profile 'test_full' on AWS batch on: - #push: pull_request: branches: - master - #- dev # just for testing purposes, to be removed release: types: [published] From 5d88e0c766c920d9324df378a0d3d1bc6cec56a3 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 27 Mar 2020 13:11:15 +0000 Subject: [PATCH 157/226] Change pull_request to pull --- .github/workflows/awstest.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/.github/workflows/awstest.yml b/.github/workflows/awstest.yml index b7a47e78..8ccdb4cf 100644 --- a/.github/workflows/awstest.yml +++ b/.github/workflows/awstest.yml @@ -3,7 +3,7 @@ name: nf-core AWS test # It runs the -profile 'test_full' on AWS batch on: - pull_request: + push: branches: - master release: From 3048f996cfac6d86dff9691324bc4b405c18f3b1 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 27 Mar 2020 15:20:50 +0000 Subject: [PATCH 158/226] Push to dev --- .github/workflows/awstest.yml | 1 + 1 file changed, 1 insertion(+) diff --git a/.github/workflows/awstest.yml b/.github/workflows/awstest.yml index 8ccdb4cf..542c1976 100644 --- a/.github/workflows/awstest.yml +++ b/.github/workflows/awstest.yml @@ -6,6 +6,7 @@ on: push: branches: - master + - dev release: types: [published] From 5f81accd28cd7eda2bb373df6fabb941ce321c15 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 27 Mar 2020 17:13:02 +0000 Subject: [PATCH 159/226] Update schema --- nextflow_schema.json | 100 +++++++++++++++++++++---------------------- 1 file changed, 49 insertions(+), 51 deletions(-) diff --git a/nextflow_schema.json b/nextflow_schema.json index 4487fd5e..83c5b73e 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -5,7 +5,7 @@ "description": "ATACSeq peak-calling and differential analysis pipeline.", "type": "object", "properties": { - "Generic options": { + "Input/output options": { "type": "object", "properties": { "input": { @@ -29,16 +29,24 @@ "fa_icon": "fas fa-align-justify", "help_text": "" }, - "fingerprint_bins": { - "type": "integer", - "default": 500000, - "description": "Number of genomic bins to use when calculating deepTools fingerprint plot.", - "fa_icon": "fas fa-dumpster" - }, "seq_center": { "type": "string", "description": "Sequencing center information to be added to read group of BAM files.", "fa_icon": "fas fa-synagogue" + }, + "outdir": { + "type": "string", + "description": "The output directory where the results will be saved.", + "default": "./results", + "fa_icon": "fas fa-folder-open", + "help_text": "" + }, + "email": { + "type": "string", + "description": "Email address for completion summary.", + "fa_icon": "fas fa-envelope", + "help_text": "An email address to send a summary email to when the pipeline is completed.", + "pattern": "^([a-zA-Z0-9_\\-\\.]+)@([a-zA-Z0-9_\\-\\.]+)\\.([a-zA-Z]{2,5})$" } }, "required": [ @@ -216,6 +224,22 @@ "description": "Save the intermediate BAM files from the alignment step.", "help_text": "By default, intermediate BAM files will not be saved. The final BAM files created after the appropriate filtering step are always saved to limit storage usage. Set this parameter to also save other intermediate BAM files.", "fa_icon": "fas fa-save" + }, + "bamtools_filter_pe_config": { + "type": "string", + "default": "$baseDir/assets/bamtools_filter_pe.json", + "hidden": true, + "description": "BAMTools JSON file with custom filters for paired-end data.", + "fa_icon": "fas fa-cog", + "help_text": "" + }, + "bamtools_filter_se_config": { + "type": "string", + "default": "$baseDir/assets/bamtools_filter_se.json", + "hidden": true, + "description": "BAMTools JSON file with custom filters for single-end data.", + "fa_icon": "fas fa-cog", + "help_text": "" } }, "fa_icon": "fas fa-map-signs" @@ -334,35 +358,6 @@ }, "fa_icon": "fas fa-forward" }, - "Pipeline config options": { - "type": "object", - "properties": { - "multiqc_config": { - "type": "string", - "description": "Custom config file to supply to MultiQC.", - "fa_icon": "fas fa-cog", - "hidden": true, - "help_text": "" - }, - "bamtools_filter_pe_config": { - "type": "string", - "default": "$baseDir/assets/bamtools_filter_pe.json", - "hidden": true, - "description": "BAMTools JSON file with custom filters for paired-end data.", - "fa_icon": "fas fa-cog", - "help_text": "" - }, - "bamtools_filter_se_config": { - "type": "string", - "default": "$baseDir/assets/bamtools_filter_se.json", - "hidden": true, - "description": "BAMTools JSON file with custom filters for single-end data.", - "fa_icon": "fas fa-cog", - "help_text": "" - } - }, - "fa_icon": "fas fa-cog" - }, "Institutional config options": { "type": "object", "properties": { @@ -443,7 +438,7 @@ }, "fa_icon": "fab fa-acquisitions-incorporated" }, - "Pipeline template options": { + "Generic options": { "type": "object", "properties": { "help": { @@ -453,12 +448,12 @@ "fa_icon": "fas fa-question-circle", "default": false }, - "outdir": { - "type": "string", - "description": "The output directory where the results will be saved.", - "default": "./results", - "fa_icon": "fas fa-folder-open", - "help_text": "" + "fingerprint_bins": { + "type": "integer", + "default": 500000, + "hidden": true, + "description": "Number of genomic bins to use when calculating deepTools fingerprint plot.", + "fa_icon": "fas fa-dumpster" }, "publish_dir_mode": { "type": "string", @@ -480,20 +475,15 @@ "type": "string", "description": "Workflow name.", "fa_icon": "fas fa-fingerprint", + "hidden": true, "help_text": "A custom name for the pipeline run. Unlike the core nextflow `-name` option with one hyphen this parameter can be reused multiple times, for example if using `-resume`. Passed through to steps such as MultiQC and used for things like report filenames and titles." }, - "email": { - "type": "string", - "description": "Email address for completion summary.", - "fa_icon": "fas fa-envelope", - "help_text": "An email address to send a summary email to when the pipeline is completed.", - "pattern": "^([a-zA-Z0-9_\\-\\.]+)@([a-zA-Z0-9_\\-\\.]+)\\.([a-zA-Z]{2,5})$" - }, "email_on_fail": { "type": "string", "description": "Email address for completion summary, only when pipeline fails.", "fa_icon": "fas fa-exclamation-triangle", "pattern": "^([a-zA-Z0-9_\\-\\.]+)@([a-zA-Z0-9_\\-\\.]+)\\.([a-zA-Z]{2,5})$", + "hidden": true, "help_text": "An email address to send a summary email to when the pipeline is completed - ONLY sent if the pipeline does not exit successfully." }, "plaintext_email": { @@ -520,6 +510,13 @@ "default": false, "help_text": "" }, + "multiqc_config": { + "type": "string", + "description": "Custom config file to supply to MultiQC.", + "fa_icon": "fas fa-cog", + "hidden": true, + "help_text": "" + }, "tracedir": { "type": "string", "description": "Directory to keep pipeline Nextflow logs and reports.", @@ -538,5 +535,6 @@ }, "fa_icon": "fas fa-file-import" } - } + }, + "required": ["Input/output options"] } From 4ee6b312be05122e70318c92f6b87efb96bdfc42 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 29 Apr 2020 17:39:55 +0100 Subject: [PATCH 160/226] import errno --- bin/bampe_rm_orphan.py | 1 + bin/get_autosomes.py | 1 + bin/igv_files_to_session.py | 1 + bin/macs2_merged_expand.py | 1 + 4 files changed, 4 insertions(+) diff --git a/bin/bampe_rm_orphan.py b/bin/bampe_rm_orphan.py index 439a53ee..5b0a6f72 100755 --- a/bin/bampe_rm_orphan.py +++ b/bin/bampe_rm_orphan.py @@ -8,6 +8,7 @@ import os import pysam +import errno import argparse ############################################ diff --git a/bin/get_autosomes.py b/bin/get_autosomes.py index a8603b6b..2d530561 100755 --- a/bin/get_autosomes.py +++ b/bin/get_autosomes.py @@ -7,6 +7,7 @@ ####################################################################### import os +import errno import argparse ############################################ diff --git a/bin/igv_files_to_session.py b/bin/igv_files_to_session.py index 6dc6aabe..48e749c8 100755 --- a/bin/igv_files_to_session.py +++ b/bin/igv_files_to_session.py @@ -7,6 +7,7 @@ ####################################################################### import os +import errno import argparse ############################################ diff --git a/bin/macs2_merged_expand.py b/bin/macs2_merged_expand.py index 55c3a057..f4e84a14 100755 --- a/bin/macs2_merged_expand.py +++ b/bin/macs2_merged_expand.py @@ -7,6 +7,7 @@ ####################################################################### import os +import errno import argparse ############################################ From d8d827d8b2b19eab0b527dd7035dd09d244f003c Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 28 May 2020 16:29:52 +0100 Subject: [PATCH 161/226] Fix #102 --- bin/featurecounts_deseq2.r | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/bin/featurecounts_deseq2.r b/bin/featurecounts_deseq2.r index e19fd977..b75335a5 100755 --- a/bin/featurecounts_deseq2.r +++ b/bin/featurecounts_deseq2.r @@ -78,7 +78,7 @@ if (file.exists(opt$outdir) == FALSE) { setwd(opt$outdir) samples.vec <- sort(colnames(count.table)) -groups <- substr(samples.vec, 1, nchar(samples.vec)-3) +groups <- sub("_[^_]+$", "", samples.vec) print(unique(groups)) if (length(unique(groups)) == 1) { quit(save = "no", status = 0, runLast = FALSE) From 60380e60fc6b84648470771b9d96761ca02813e2 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 28 May 2020 16:46:25 +0100 Subject: [PATCH 162/226] Update CHANGELOG --- CHANGELOG.md | 1 + 1 file changed, 1 insertion(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index d23a798a..bc4856d1 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -26,6 +26,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * [nf-core/chipseq#132](https://github.com/nf-core/chipseq/issues/132) - BigWig Error: sort: cannot create temporary file in '': Read-only file system * [#73](https://github.com/nf-core/atacseq/issues/73) - macs_annotatePeaks.mLb.clN.summary.txt file is not created * [#86](https://github.com/nf-core/atacseq/issues/86) - bug in the plot_homer_annotatepeaks.r script +* [#102](https://github.com/nf-core/atacseq/issues/102) - Incorrect Group ID assigned by featurecounts_deseq2.r * Make executables in `bin/` compatible with Python 3 ### `Dependencies` From 61bdc50a6d7cf4ca91be47c526da7360d1db6257 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 28 May 2020 17:57:02 +0100 Subject: [PATCH 163/226] Revert bump-versions --- .github/workflows/ci.yml | 4 ++-- Dockerfile | 4 ++-- environment.yml | 2 +- nextflow.config | 4 ++-- 4 files changed, 7 insertions(+), 7 deletions(-) diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml index a3cff843..baa82785 100644 --- a/.github/workflows/ci.yml +++ b/.github/workflows/ci.yml @@ -22,7 +22,7 @@ jobs: - name: Pull docker image run: | docker pull nfcore/atacseq:dev - docker tag nfcore/atacseq:dev nfcore/atacseq:1.2.0 + docker tag nfcore/atacseq:dev nfcore/atacseq:dev - name: Run pipeline with test data run: | nextflow run ${GITHUB_WORKSPACE} -profile test,docker @@ -44,7 +44,7 @@ jobs: - name: Pull docker image run: | docker pull nfcore/atacseq:dev - docker tag nfcore/atacseq:dev nfcore/atacseq:1.2.0 + docker tag nfcore/atacseq:dev nfcore/atacseq:dev - name: Run pipeline with various options run: | nextflow run ${GITHUB_WORKSPACE} -profile test,docker ${{ matrix.parameters }} diff --git a/Dockerfile b/Dockerfile index 715e6892..ecae8934 100644 --- a/Dockerfile +++ b/Dockerfile @@ -7,10 +7,10 @@ COPY environment.yml / RUN conda env create -f /environment.yml && conda clean -a # Add conda installation dir to PATH (instead of doing 'conda activate') -ENV PATH /opt/conda/envs/nf-core-atacseq-1.2.0/bin:$PATH +ENV PATH /opt/conda/envs/nf-core-atacseq-1.1.1dev/bin:$PATH # Dump the details of the installed packages to a file for posterity -RUN conda env export --name nf-core-atacseq-1.2.0 > nf-core-atacseq-1.2.0.yml +RUN conda env export --name nf-core-atacseq-1.1.1dev > nf-core-atacseq-1.1.1dev.yml # Instruct R processes to use these empty files instead of clashing with a local version RUN touch .Rprofile diff --git a/environment.yml b/environment.yml index 535b7768..af38f938 100755 --- a/environment.yml +++ b/environment.yml @@ -1,6 +1,6 @@ # You can use this file to create a conda environment for this pipeline: # conda env create -f environment.yml -name: nf-core-atacseq-1.2.0 +name: nf-core-atacseq-1.1.1dev channels: - conda-forge - bioconda diff --git a/nextflow.config b/nextflow.config index 2d5009de..7e00b824 100644 --- a/nextflow.config +++ b/nextflow.config @@ -94,7 +94,7 @@ params { // Container slug. Stable releases should specify release tag! // Developmental code should specify :dev -process.container = 'nfcore/atacseq:1.2.0' +process.container = 'nfcore/atacseq:dev' // Load base.config by default for all pipelines includeConfig 'conf/base.config' @@ -167,7 +167,7 @@ manifest { description = 'ATACSeq peak-calling and differential analysis pipeline.' mainScript = 'main.nf' nextflowVersion = '>=19.10.0' - version = '1.2.0' + version = '1.1.1dev' } // Function to ensure that resource requirements don't go beyond From ad0a7d5c64c0be14982fca9053fa85faa9821463 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 28 May 2020 17:57:15 +0100 Subject: [PATCH 164/226] Update CHANGELOG --- CHANGELOG.md | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index bc4856d1..4a9e34af 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -22,11 +22,11 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Fixed` -* [nf-core/chipseq#118](https://github.com/nf-core/chipseq/issues/118) - Running on with SGE -* [nf-core/chipseq#132](https://github.com/nf-core/chipseq/issues/132) - BigWig Error: sort: cannot create temporary file in '': Read-only file system * [#73](https://github.com/nf-core/atacseq/issues/73) - macs_annotatePeaks.mLb.clN.summary.txt file is not created * [#86](https://github.com/nf-core/atacseq/issues/86) - bug in the plot_homer_annotatepeaks.r script * [#102](https://github.com/nf-core/atacseq/issues/102) - Incorrect Group ID assigned by featurecounts_deseq2.r +* [nf-core/chipseq#118](https://github.com/nf-core/chipseq/issues/118) - Running on with SGE +* [nf-core/chipseq#132](https://github.com/nf-core/chipseq/issues/132) - BigWig Error: sort: cannot create temporary file in '': Read-only file system * Make executables in `bin/` compatible with Python 3 ### `Dependencies` From 3d6f33f49c0c57e465b3a5318cfe109c9c1e62e1 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 28 May 2020 17:57:21 +0100 Subject: [PATCH 165/226] Update CITATIOSN --- CITATIONS.md | 11 ++++++++--- 1 file changed, 8 insertions(+), 3 deletions(-) diff --git a/CITATIONS.md b/CITATIONS.md index 2ef37fed..9af4aba2 100644 --- a/CITATIONS.md +++ b/CITATIONS.md @@ -1,9 +1,14 @@ # nf-core/atacseq: Citations -## Pipeline tools +## [nf-core](https://www.ncbi.nlm.nih.gov/pubmed/32055031/) + +> Ewels PA, Peltzer A, Fillinger S, Patel H, Alneberg J, Wilm A, Garcia MU, Di Tommaso P, Nahnsen S. The nf-core framework for community-curated bioinformatics pipelines. Nat Biotechnol. 2020 Mar;38(3):276-278. doi: 10.1038/s41587-020-0439-x. PubMed PMID: 32055031. ReadCube: [Full Access Link](https://rdcu.be/b1GjZ). + +## [Nextflow](https://www.ncbi.nlm.nih.gov/pubmed/28398311/) -* [Nextflow](https://www.ncbi.nlm.nih.gov/pubmed/28398311/) - > Di Tommaso P, Chatzou M, Floden EW, Barja PP, Palumbo E, Notredame C. Nextflow enables reproducible computational workflows. Nat Biotechnol. 2017 Apr 11;35(4):316-319. doi: 10.1038/nbt.3820. PubMed PMID: 28398311. +> Di Tommaso P, Chatzou M, Floden EW, Barja PP, Palumbo E, Notredame C. Nextflow enables reproducible computational workflows. Nat Biotechnol. 2017 Apr 11;35(4):316-319. doi: 10.1038/nbt.3820. PubMed PMID: 28398311. + +## Pipeline tools * [BWA](https://www.ncbi.nlm.nih.gov/pubmed/19451168/) > Li H, Durbin R. Fast and accurate short read alignment with Burrows-Wheeler transform. Bioinformatics. 2009 Jul 15;25(14):1754-60. doi: 10.1093/bioinformatics/btp324. Epub 2009 May 18. PubMed PMID: 19451168; PubMed Central PMCID: PMC2705234. From 5a20b8f5f3084c15d9e67f27e587319fdc519d6f Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 28 May 2020 18:41:51 +0100 Subject: [PATCH 166/226] Update package versions --- CHANGELOG.md | 31 ++++++++++++++++--------------- 1 file changed, 16 insertions(+), 15 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 4a9e34af..ecaf3995 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -32,30 +32,31 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Dependencies` * Add python `3.7.6` -* Add markdown `3.1.1` -* Add pymdown-extensions `6.0` -* Add pygments `2.5.2` +* Add markdown `3.2.2` +* Add pymdown-extensions `7.1` +* Add pygments `2.6.1` * Add pigz `2.3.4` -* Add r-tidyr `1.0.2` +* Add r-tidyr `1.1.0` * Add bioconductor-biocparallel `1.20.0` -* Update gawk `4.2.1` -> `5.0.1` +* Update gawk `4.2.1` -> `5.1.0` * Update r-base `3.4.1` -> `3.6.2` -* Update r-optparse `1.6.0` -> `1.6.4` -* Update r-ggplot2 `3.1.0` -> `3.2.1` +* Update r-optparse `1.6.0` -> `1.6.6` +* Update r-ggplot2 `3.1.0` -> `3.3.0` * Update r-pheatmap `1.0.10` -> `1.0.12` -* Update r-lattice `0.20_35` -> `0.20_40` +* Update r-lattice `0.20_35` -> `0.20_41` * Update r-upsetr `1.3.3` -> `1.4.0` -* Update r-scales `1.0.0` -> `1.1.0` -* Update r-xfun `0.3` -> `0.12` +* Update r-scales `1.0.0` -> `1.1.1` +* Update r-xfun `0.3` -> `0.14` * Update fastqc `0.11.8` -> `0.11.9` * Update trim-galore `0.5.0` -> `0.6.5` -* Update picard `2.19.0` -> `2.21.9` -* Update pysam `0.15.2` -> `0.15.4` +* Update samtools `1.9` -> `1.10` +* Update picard `2.19.0` -> `2.22.8` +* Update pysam `0.15.2` -> `0.15.3` * Update bedtools `2.27.1` -> `2.29.2` * Update ucsc-bedgraphtobigwig `377` -> `357` -* Update deeptools `3.2.1` -> `3.3.2` -* Update macs2 `2.1.2` -> `2.2.6` -* Update homer `4.9.1` -> `4.10` +* Update deeptools `3.2.1` -> `3.4.3` +* Update macs2 `2.1.2` -> `2.2.7.1` +* Update homer `4.9.1` -> `4.11` * Update ataqv `1.0.0` -> `1.1.1` * Update subread `1.6.4` -> `2.0.0` * Update multiqc `1.7` -> `1.8` From e69998b8db9e7155011e58d4cfb0c6bae4932758 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 28 May 2020 18:41:59 +0100 Subject: [PATCH 167/226] Update environment --- environment.yml | 32 ++++++++++++++++---------------- 1 file changed, 16 insertions(+), 16 deletions(-) diff --git a/environment.yml b/environment.yml index af38f938..ea559a1c 100755 --- a/environment.yml +++ b/environment.yml @@ -8,35 +8,35 @@ channels: dependencies: ## conda-forge packages - conda-forge::python=3.7.6 - - conda-forge::markdown=3.1.1 - - conda-forge::pymdown-extensions=6.0 - - conda-forge::pygments=2.5.2 + - conda-forge::markdown=3.2.2 + - conda-forge::pymdown-extensions=7.1 + - conda-forge::pygments=2.6.1 - conda-forge::r-base=3.6.2 - - conda-forge::r-optparse=1.6.4 + - conda-forge::r-optparse=1.6.6 - conda-forge::r-rcolorbrewer=1.1_2 - - conda-forge::r-ggplot2=3.2.1 - - conda-forge::r-tidyr=1.0.2 - - conda-forge::r-scales=1.1.0 + - conda-forge::r-ggplot2=3.3.0 + - conda-forge::r-tidyr=1.1.0 + - conda-forge::r-scales=1.1.1 - conda-forge::r-pheatmap=1.0.12 - - conda-forge::r-lattice=0.20_40 + - conda-forge::r-lattice=0.20_41 - conda-forge::r-upsetr=1.4.0 - - conda-forge::r-xfun=0.12 - - conda-forge::gawk=5.0.1 + - conda-forge::r-xfun=0.14 + - conda-forge::gawk=5.1.0 - conda-forge::pigz=2.3.4 ## Required for TrimGalore multi-threading ## bioconda packages - bioconda::fastqc=0.11.9 - bioconda::trim-galore=0.6.5 - bioconda::bwa=0.7.17 - - bioconda::samtools=1.9 - - bioconda::picard=2.21.9 + - bioconda::samtools=1.10 + - bioconda::picard=2.22.8 - bioconda::bamtools=2.5.1 - - bioconda::pysam=0.15.4 + - bioconda::pysam=0.15.3 - bioconda::bedtools=2.29.2 - bioconda::ucsc-bedgraphtobigwig=357 - - bioconda::deeptools=3.3.2 - - bioconda::macs2=2.2.6 - - bioconda::homer=4.10 + - bioconda::deeptools=3.4.3 + - bioconda::macs2=2.2.7.1 + - bioconda::homer=4.11 - bioconda::ataqv=1.1.1 - bioconda::subread=2.0.0 - bioconda::preseq=2.0.3 From b21264bc90f2b6e5ba90e1c86c5159405aedf703 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 28 May 2020 21:46:12 +0100 Subject: [PATCH 168/226] Update CHANGELOG --- CHANGELOG.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index ecaf3995..e8b4dc51 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -3,7 +3,7 @@ The format is based on [Keep a Changelog](http://keepachangelog.com/en/1.0.0/) and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0.html). -## [1.2.0] - 2020-03-20 +## [1.2.0] - 2020-06-04 ### `Added` From 5200e75e2523dbd0a300984103e3529852555158 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 28 May 2020 22:00:33 +0100 Subject: [PATCH 169/226] Remove CI test --- .github/workflows/ci.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml index baa82785..86b5670e 100644 --- a/.github/workflows/ci.yml +++ b/.github/workflows/ci.yml @@ -34,7 +34,7 @@ jobs: runs-on: ubuntu-latest strategy: matrix: - parameters: [--single_end, --skip_trimming, --skip_merge_replicates, --skip_consensus_peaks, --skip_multiqc] + parameters: [--single_end, --skip_trimming, --skip_merge_replicates, --skip_consensus_peaks] steps: - uses: actions/checkout@v2 - name: Install Nextflow From f7be7694d8f2d4f2b1be6969982127a2dfe68102 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 29 May 2020 12:43:17 +0100 Subject: [PATCH 170/226] Remove AWS test --- .github/workflows/awstest.yml | 1 - 1 file changed, 1 deletion(-) diff --git a/.github/workflows/awstest.yml b/.github/workflows/awstest.yml index 542c1976..8ccdb4cf 100644 --- a/.github/workflows/awstest.yml +++ b/.github/workflows/awstest.yml @@ -6,7 +6,6 @@ on: push: branches: - master - - dev release: types: [published] From b2c1a517fc12ff6af978a6879dd047160ec005c8 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 29 May 2020 12:43:23 +0100 Subject: [PATCH 171/226] Update CHANGELOG --- CHANGELOG.md | 1 + 1 file changed, 1 insertion(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index e8b4dc51..ecf512d1 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -37,6 +37,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * Add pygments `2.6.1` * Add pigz `2.3.4` * Add r-tidyr `1.1.0` +* Add r-reshape2 `1.4.4` * Add bioconductor-biocparallel `1.20.0` * Update gawk `4.2.1` -> `5.1.0` * Update r-base `3.4.1` -> `3.6.2` From aa7f4c564767b7a4bca493fe9856b87bd13ff67f Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 29 May 2020 12:43:32 +0100 Subject: [PATCH 172/226] Add reshape2 --- environment.yml | 1 + 1 file changed, 1 insertion(+) diff --git a/environment.yml b/environment.yml index ea559a1c..2c7bab71 100755 --- a/environment.yml +++ b/environment.yml @@ -14,6 +14,7 @@ dependencies: - conda-forge::r-base=3.6.2 - conda-forge::r-optparse=1.6.6 - conda-forge::r-rcolorbrewer=1.1_2 + - conda-forge::r-reshape2=1.4.4 - conda-forge::r-ggplot2=3.3.0 - conda-forge::r-tidyr=1.1.0 - conda-forge::r-scales=1.1.1 From da3cb03f54bea0e4a3aa8024b53c60629418df6f Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 29 May 2020 12:43:41 +0100 Subject: [PATCH 173/226] Change process names --- main.nf | 72 ++++++++++++++++++++++++++++----------------------------- 1 file changed, 36 insertions(+), 36 deletions(-) diff --git a/main.nf b/main.nf index cba830ce..516acda4 100644 --- a/main.nf +++ b/main.nf @@ -318,7 +318,7 @@ if (!params.macs_gsize) { /* * PREPROCESSING: Reformat design file and check validitiy */ -process CheckDesign { +process CHECK_DESIGN { tag "$design" publishDir "${params.outdir}/pipeline_info", mode: params.publish_dir_mode @@ -382,7 +382,7 @@ multipleGroups = design_multiple_samples * PREPROCESSING: Build BWA index */ if (!params.bwa_index) { - process BWAIndex { + process BWA_INDEX { tag "$fasta" label 'process_high' publishDir path: { params.save_reference ? "${params.outdir}/genome" : params.outdir }, @@ -406,7 +406,7 @@ if (!params.bwa_index) { * PREPROCESSING: Generate gene BED file */ if (!params.gene_bed) { - process MakeGeneBED { + process MAKE_GENE_BED { tag "$gtf" label 'process_low' publishDir "${params.outdir}/genome", mode: params.publish_dir_mode @@ -428,7 +428,7 @@ if (!params.gene_bed) { * PREPROCESSING: Generate TSS BED file */ if (!params.tss_bed) { - process MakeTSSBED { + process MAKE_TSS_BED { tag "$bed" publishDir "${params.outdir}/genome", mode: params.publish_dir_mode @@ -448,7 +448,7 @@ if (!params.tss_bed) { /* * PREPROCESSING: Prepare genome intervals for filtering */ -process MakeGenomeFilter { +process MAKE_GENOME_FILTER { tag "$fasta" publishDir "${params.outdir}/genome", mode: params.publish_dir_mode @@ -487,7 +487,7 @@ process MakeGenomeFilter { /* * STEP 1: FastQC */ -process FastQC { +process FASTQC { tag "$name" label 'process_medium' publishDir "${params.outdir}/fastqc", mode: params.publish_dir_mode, @@ -537,7 +537,7 @@ if (params.skip_trimming) { ch_trimgalore_results_mqc = Channel.empty() ch_trimgalore_fastqc_reports_mqc = Channel.empty() } else { - process TrimGalore { + process TRIMGALORE { tag "$name" label 'process_high' publishDir "${params.outdir}/trim_galore", mode: params.publish_dir_mode, @@ -602,7 +602,7 @@ if (params.skip_trimming) { /* * STEP 3.1: Map read(s) with bwa mem */ -process BWAMem { +process BWA_MEM { tag "$name" label 'process_high' @@ -633,7 +633,7 @@ process BWAMem { /* * STEP 3.2: Convert BAM to coordinate sorted BAM */ -process SortBAM { +process SORT_BAM { tag "$name" label 'process_medium' if (params.save_align_intermeds) { @@ -681,7 +681,7 @@ ch_sort_bam_merge .map { it -> [ it[0], it[1].flatten() ] } .set { ch_sort_bam_merge } -process MergedLibBAM { +process MERGED_LIB_BAM { tag "$name" label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary", mode: params.publish_dir_mode, @@ -758,7 +758,7 @@ process MergedLibBAM { /* * STEP 4.2: Filter BAM file at merged library-level */ -process MergedLibBAMFilter { +process MERGED_LIB_BAM_FILTER { tag "$name" label 'process_medium' publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: params.publish_dir_mode, @@ -831,7 +831,7 @@ if (params.single_end) { ch_mlib_filter_bam_stats_mqc .set { ch_mlib_rm_orphan_stats_mqc } } else { - process MergedLibBAMRemoveOrphan { + process MERGED_LIB_BAM_REMOVE_ORPHAN { tag "$name" label 'process_medium' publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: params.publish_dir_mode, @@ -885,7 +885,7 @@ if (params.single_end) { /* * STEP 5.1: Preseq analysis after merging libraries and before filtering */ -process MergedLibPreseq { +process MERGED_LIB_PRESEQ { tag "$name" label 'process_low' publishDir "${params.outdir}/bwa/mergedLibrary/preseq", mode: params.publish_dir_mode @@ -918,7 +918,7 @@ process MergedLibPreseq { /* * STEP 5.2: Picard CollectMultipleMetrics after merging libraries and filtering */ -process MergedLibMetrics { +process MERGED_LIB_PICARD_METRICS { tag "$name" label 'process_medium' publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: params.publish_dir_mode, @@ -960,7 +960,7 @@ process MergedLibMetrics { /* * STEP 5.3: Read depth normalised bigWig */ -process MergedLibBigWig { +process MERGED_LIB_BIGWIG { tag "$name" label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/bigwig", mode: params.publish_dir_mode, @@ -997,7 +997,7 @@ process MergedLibBigWig { /* * STEP 5.4: Generate gene body coverage plot with deepTools plotProfile */ -process MergedLibPlotProfile { +process MERGED_LIB_PLOTPROFILE { tag "$name" label 'process_high' publishDir "${params.outdir}/bwa/mergedLibrary/deepTools/plotProfile", mode: params.publish_dir_mode @@ -1037,7 +1037,7 @@ process MergedLibPlotProfile { /* * STEP 5.5: deepTools plotFingerprint */ -process MergedLibPlotFingerprint { +process MERGED_LIB_PLOTFINGERPRINT { tag "$name" label 'process_high' publishDir "${params.outdir}/bwa/mergedLibrary/deepTools/plotFingerprint", mode: params.publish_dir_mode @@ -1080,7 +1080,7 @@ process MergedLibPlotFingerprint { /* * STEP 6.1: Call peaks with MACS2 and calculate FRiP score */ -process MergedLibMACSCallPeak { +process MERGED_LIB_MACS2 { tag "$name" label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}", mode: params.publish_dir_mode, @@ -1135,7 +1135,7 @@ process MergedLibMACSCallPeak { /* * STEP 6.2: Annotate peaks with HOMER */ -process MergedLibAnnotatePeaks { +process MERGED_LIB_MACS2_ANNOTATE { tag "$name" label "process_medium" publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}", mode: params.publish_dir_mode @@ -1167,7 +1167,7 @@ process MergedLibAnnotatePeaks { /* * STEP 6.3: Aggregated QC plots for peaks, FRiP and peak-to-gene annotation */ -process MergedLibPeakQC { +process MERGED_LIB_MACS2_QC { label "process_medium" publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/qc", mode: params.publish_dir_mode @@ -1205,7 +1205,7 @@ process MergedLibPeakQC { /* * STEP 6.4: Consensus peaks across samples, create boolean filtering file, SAF file for featureCounts and UpSetR plot for intersection */ -process MergedLibConsensusPeakSet { +process MERGED_LIB_CONSENSUS { label 'process_long' publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: params.publish_dir_mode, saveAs: { filename -> @@ -1258,7 +1258,7 @@ process MergedLibConsensusPeakSet { /* * STEP 6.5: Annotate consensus peaks with HOMER, and add annotation to boolean output file */ -process MergedLibConsensusPeakSetAnnotate { +process MERGED_LIB_CONSENSUS_ANNOTATE { label "process_medium" publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: params.publish_dir_mode @@ -1293,7 +1293,7 @@ process MergedLibConsensusPeakSetAnnotate { /* * STEP 6.6: Count reads in consensus peaks with featureCounts */ -process MergedLibConsensusPeakSetCounts { +process MERGED_LIB_CONSENSUS_COUNTS { label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: params.publish_dir_mode @@ -1328,7 +1328,7 @@ process MergedLibConsensusPeakSetCounts { /* * STEP 6.7: Differential analysis with DESeq2 */ -process MergedLibConsensusPeakSetDESeq { +process MERGED_LIB_CONSENSUS_DESEQ2 { label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus/deseq2", mode: params.publish_dir_mode, saveAs: { filename -> @@ -1376,7 +1376,7 @@ process MergedLibConsensusPeakSetDESeq { /* * STEP 6.8: Run ataqv on BAM file and corresponding peaks */ -process MergedLibAtaqv { +process MERGED_LIB_ATAQV { tag "$name" label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/ataqv/${PEAK_TYPE}", mode: params.publish_dir_mode @@ -1414,7 +1414,7 @@ process MergedLibAtaqv { /* * STEP 6.9: Run ataqv mkarv on all JSON files to render web app */ -process MergedLibAtaqvMkarv { +process MERGED_LIB_ATAQV_MKARV { label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/ataqv/${PEAK_TYPE}", mode: params.publish_dir_mode @@ -1454,7 +1454,7 @@ ch_mlib_rm_orphan_bam_mrep .map { it -> [ it[0], it[1].flatten() ] } .set { ch_mlib_rm_orphan_bam_mrep } -process MergedRepBAM { +process MERGED_REP_BAM { tag "$name" label 'process_medium' publishDir "${params.outdir}/bwa/mergedReplicate", mode: params.publish_dir_mode, @@ -1537,7 +1537,7 @@ process MergedRepBAM { /* * STEP 8.1: Read depth normalised bigWig */ -process MergedRepBigWig { +process MERGED_REP_BIGWIG { tag "$name" label 'process_medium' publishDir "${params.outdir}/bwa/mergedReplicate/bigwig", mode: params.publish_dir_mode, @@ -1577,7 +1577,7 @@ process MergedRepBigWig { /* * STEP 8.2: Call peaks with MACS2 and calculate FRiP score */ -process MergedRepMACSCallPeak { +process MERGED_REP_MACS2 { tag "$name" label 'process_medium' publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}", mode: params.publish_dir_mode, @@ -1631,7 +1631,7 @@ process MergedRepMACSCallPeak { /* * STEP 8.3: Annotate peaks with HOMER */ -process MergedRepAnnotatePeaks { +process MERGED_REP_MACS2_ANNOTATE { tag "$name" label "process_medium" publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}", mode: params.publish_dir_mode @@ -1663,7 +1663,7 @@ process MergedRepAnnotatePeaks { /* * STEP 8.4: Aggregated QC plots for peaks, FRiP and peak-to-gene annotation */ -process MergedRepPeakQC { +process MERGED_REP_MACS2_QC { label "process_medium" publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/qc", mode: params.publish_dir_mode @@ -1701,7 +1701,7 @@ process MergedRepPeakQC { /* * STEP 8.5: Consensus peaks across samples, create boolean filtering file, SAF file for featureCounts and UpSetR plot for intersection */ -process MergedRepConsensusPeakSet { +process MERGED_REP_CONSENSUS { label 'process_long' publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: params.publish_dir_mode, saveAs: { filename -> @@ -1753,7 +1753,7 @@ process MergedRepConsensusPeakSet { /* * STEP 8.6: Annotate consensus peaks with HOMER, and add annotation to boolean output file */ -process MergedRepConsensusPeakSetAnnotate { +process MERGED_REP_CONSENSUS_ANNOTATE { label "process_medium" publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: params.publish_dir_mode @@ -1788,7 +1788,7 @@ process MergedRepConsensusPeakSetAnnotate { /* * STEP 8.7: Count reads in consensus peaks with featureCounts */ -process MergedRepConsensusPeakSetCounts { +process MERGED_REP_CONSENSUS_COUNTS { label 'process_medium' publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: params.publish_dir_mode @@ -1823,7 +1823,7 @@ process MergedRepConsensusPeakSetCounts { /* * STEP 8.8: Differential analysis with DESeq2 */ -process MergedRepConsensusPeakSetDESeq { +process MERGED_REP_CONSENSUS_DESEQ2 { label 'process_medium' publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus/deseq2", mode: params.publish_dir_mode, saveAs: { filename -> @@ -1973,7 +1973,7 @@ Channel.from(summary.collect{ [it.key, it.value] }) /* * STEP 10: MultiQC */ -process MultiQC { +process MULTIQC { publishDir "${params.outdir}/multiqc/${PEAK_TYPE}", mode: params.publish_dir_mode when: From 894857bb074ac052394386dc5d1f956bb310aff3 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 29 May 2020 13:29:52 +0100 Subject: [PATCH 174/226] Update MultiQC config --- assets/multiqc_config.yaml | 41 ++++++++++++++++++++++++-------------- main.nf | 4 +--- 2 files changed, 27 insertions(+), 18 deletions(-) diff --git a/assets/multiqc_config.yaml b/assets/multiqc_config.yaml index e76c08b7..5d07c929 100755 --- a/assets/multiqc_config.yaml +++ b/assets/multiqc_config.yaml @@ -3,19 +3,20 @@ report_comment: > analysis pipeline. For information about how to interpret these results, please see the documentation. -export_plots: true +data_format: 'yaml' -fn_clean_exts: - - 'fastq.gz' - - '_trimmed' - - '_val' - - 'sorted.bam' - - '.Lb' - - 'mkD' - - 'clN' - - 'mLb' - - 'mRp' - - '_peaks' +run_modules: + - custom_content + - fastqc + - cutadapt + - samtools + - picard + - preseq + - featureCounts + - deeptools + +exclude_modules: + - 'general_stats' module_order: - fastqc: @@ -115,12 +116,22 @@ report_section_order: order: -2100 custom_plot_config: - picard-insertsize: + picard_insert_size: cpswitch_c_active: False smooth_points: 1000 - featurecounts: cpswitch_c_active: False - featurecounts-1: cpswitch_c_active: False + +extra_fn_clean_exts: + - 'fastq.gz' + - '_trimmed' + - '_val' + - 'sorted.bam' + - '.Lb' + - 'mkD' + - 'clN' + - 'mLb' + - 'mRp' + - '_peaks' diff --git a/main.nf b/main.nf index 516acda4..5840803e 100644 --- a/main.nf +++ b/main.nf @@ -2016,15 +2016,13 @@ process MULTIQC { output: file "*multiqc_report.html" into ch_multiqc_report file "*_data" - file "multiqc_plots" script: rtitle = custom_runName ? "--title \"$custom_runName\"" : '' rfilename = custom_runName ? "--filename " + custom_runName.replaceAll('\\W','_').replaceAll('_+','_') + "_multiqc_report" : '' custom_config_file = params.multiqc_config ? "--config $mqc_custom_config" : '' """ - multiqc . -f $rtitle $rfilename $custom_config_file \\ - -m custom_content -m fastqc -m cutadapt -m samtools -m picard -m preseq -m featureCounts -m deeptools + multiqc . -f $rtitle $rfilename $custom_config_file """ } From 2c1ab3a4dbe5d2072570d7f72adb2e60f93444a9 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 29 May 2020 16:59:05 +0100 Subject: [PATCH 175/226] Add label error_ignore --- conf/base.config | 3 +++ 1 file changed, 3 insertions(+) diff --git a/conf/base.config b/conf/base.config index 92a48f32..0fab975e 100755 --- a/conf/base.config +++ b/conf/base.config @@ -38,6 +38,9 @@ process { withLabel:process_long { time = { check_max( 20.h * task.attempt, 'time' ) } } + withLabel:error_ignore { + errorStrategy = 'ignore' + } withName:get_software_versions { cache = false } From fc484e1dc9858c5bc1f8f5481ec1966ab5d0eca4 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 29 May 2020 16:59:23 +0100 Subject: [PATCH 176/226] Ignore Preseq failures --- main.nf | 1 + 1 file changed, 1 insertion(+) diff --git a/main.nf b/main.nf index 5840803e..c673f201 100644 --- a/main.nf +++ b/main.nf @@ -888,6 +888,7 @@ if (params.single_end) { process MERGED_LIB_PRESEQ { tag "$name" label 'process_low' + label 'error_ignore' publishDir "${params.outdir}/bwa/mergedLibrary/preseq", mode: params.publish_dir_mode when: From 852944907d13115aa64fb56a4e16bfb040dafa1f Mon Sep 17 00:00:00 2001 From: Harshil Patel Date: Sat, 30 May 2020 11:52:37 +0100 Subject: [PATCH 177/226] Update environment.yml --- environment.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/environment.yml b/environment.yml index 2c7bab71..66ce39c7 100755 --- a/environment.yml +++ b/environment.yml @@ -41,7 +41,7 @@ dependencies: - bioconda::ataqv=1.1.1 - bioconda::subread=2.0.0 - bioconda::preseq=2.0.3 - - bioconda::multiqc=1.8 + - bioconda::multiqc=1.9 - bioconda::bioconductor-biocparallel=1.20.0 - bioconda::bioconductor-deseq2=1.26.0 - bioconda::bioconductor-vsn=3.54.0 From 6eedb07fb79050799bd9c22bc6a448f0045535ea Mon Sep 17 00:00:00 2001 From: drpatelh Date: Mon, 1 Jun 2020 22:06:35 +0100 Subject: [PATCH 178/226] Add --skip_peak_qc parameter --- CHANGELOG.md | 2 ++ docs/usage.md | 5 +++++ main.nf | 10 ++++++---- nextflow.config | 1 + 4 files changed, 14 insertions(+), 4 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index ecf512d1..c147c815 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -15,7 +15,9 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * [#80](https://github.com/nf-core/atacseq/pull/80) - Added social preview image * [#97](https://github.com/nf-core/atacseq/issues/97) - PBC1, PBC2 from pipeline? * Update template to tools `1.9` +* Capitalise process names * Parameters: + * `--skip_peak_qc` to skip MACS2 peak QC plot generation * `--skip_consensus_peaks` to skip consensus peak generation * `--deseq2_vst` to use variance stabilizing transformation (VST) instead of regularized log transformation (rlog) with DESeq2 * `--publish_dir_mode` to customise method of publishing results to output directory [nf-core/tools#585](https://github.com/nf-core/tools/issues/585) diff --git a/docs/usage.md b/docs/usage.md index c5f0d7a8..ec64225c 100755 --- a/docs/usage.md +++ b/docs/usage.md @@ -41,6 +41,7 @@ * [`--broad_cutoff`](#--broad_cutoff) * [`--min_reps_consensus`](#--min_reps_consensus) * [`--save_macs_pileup`](#--save_macs_pileup) + * [`--skip_peak_qc`](#--skip_peak_qc) * [`--skip_consensus_peaks`](#--skip_consensus_peaks) * [Differential analysis](#differential_analysis) * [`--deseq2_vst`](#--deseq2_vst) @@ -398,6 +399,10 @@ Number of biological replicates required from a given condition for a peak to co Instruct MACS2 to create bedGraph files using the `-B --SPMR` parameters. +### `--skip_peak_qc` + +Skip MACS2 peak QC plot generation. + ### `--skip_consensus_peaks` Skip consensus peak generation, annotation and counting. diff --git a/main.nf b/main.nf index c673f201..a475fc9b 100644 --- a/main.nf +++ b/main.nf @@ -62,6 +62,7 @@ def helpMessage() { --broad_cutoff [float] Specifies broad cutoff value for MACS2. Only used when --narrow_peak isnt specified (Default: 0.1) --min_reps_consensus [int] Number of biological replicates required from a given condition for a peak to contribute to a consensus peak (Default: 1) --save_macs_pileup [bool] Instruct MACS2 to create bedGraph files normalised to signal per million reads + --skip_peak_qc [bool] Skip MACS2 peak QC plot generation --skip_consensus_peaks [bool] Skip consensus peak generation Differential analysis @@ -259,6 +260,7 @@ if (params.save_trimmed) summary['Save Trimmed'] = 'Yes' if (params.save_align_intermeds) summary['Save Intermeds'] = 'Yes' if (params.save_macs_pileup) summary['Save MACS2 Pileup'] = 'Yes' if (params.skip_merge_replicates) summary['Skip Merge Replicates'] = 'Yes' +if (params.skip_peak_qc) summary['Skip MACS2 Peak QC'] = 'Yes' if (params.skip_consensus_peaks) summary['Skip Consensus Peaks'] = 'Yes' if (params.deseq2_vst) summary['Use DESeq2 vst Transform'] = 'Yes' if (params.skip_diff_analysis) summary['Skip Differential Analysis'] = 'Yes' @@ -1173,7 +1175,7 @@ process MERGED_LIB_MACS2_QC { publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/qc", mode: params.publish_dir_mode when: - params.macs_gsize + params.macs_gsize && !params.skip_peak_qc input: file peaks from ch_mlib_macs_qc.collect{ it[1] } @@ -1669,7 +1671,7 @@ process MERGED_REP_MACS2_QC { publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/qc", mode: params.publish_dir_mode when: - !params.skip_merge_replicates && replicatesExist && params.macs_gsize + !params.skip_merge_replicates && replicatesExist && params.macs_gsize && !params.skip_peak_qc input: file peaks from ch_mrep_macs_qc.collect{ it[1] } @@ -1677,9 +1679,9 @@ process MERGED_REP_MACS2_QC { file mrep_peak_annotation_header from ch_mrep_peak_annotation_header output: - file "*.{txt,pdf}" into ch_mrep_peak_qc file "*.tsv" into ch_mrep_peak_qc_mqc - + file "*.{txt,pdf}" + script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ suffix = 'mRp.clN' """ diff --git a/nextflow.config b/nextflow.config index 7e00b824..e0adef1b 100644 --- a/nextflow.config +++ b/nextflow.config @@ -41,6 +41,7 @@ params { broad_cutoff = 0.1 min_reps_consensus = 1 save_macs_pileup = false + skip_peak_qc = false skip_consensus_peaks = false // Options: Differential analysis From 61ace7de72ccc66ab7639849dcbd076fb7e227a2 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Mon, 1 Jun 2020 22:22:40 +0100 Subject: [PATCH 179/226] Remove orphan channels --- main.nf | 64 ++++++++++++++++++++++++++++----------------------------- 1 file changed, 32 insertions(+), 32 deletions(-) diff --git a/main.nf b/main.nf index a475fc9b..304ac497 100644 --- a/main.nf +++ b/main.nf @@ -171,11 +171,11 @@ ch_mrep_deseq2_clustering_header = file("$baseDir/assets/multiqc/mrep_deseq2_clu /* -- VALIDATE INPUTS -- */ //////////////////////////////////////////////////// -if (params.input) { ch_input = file(params.input, checkIfExists: true) } else { exit 1, "Samples design file not specified!" } -if (params.gtf) { ch_gtf = file(params.gtf, checkIfExists: true) } else { exit 1, "GTF annotation file not specified!" } -if (params.gene_bed) { ch_gene_bed = file(params.gene_bed, checkIfExists: true) } -if (params.tss_bed) { ch_tss_bed = file(params.tss_bed, checkIfExists: true) } -if (params.blacklist) { ch_blacklist = Channel.fromPath(params.blacklist, checkIfExists: true) } else { ch_blacklist = Channel.empty() } +if (params.input) { ch_input = file(params.input, checkIfExists: true) } else { exit 1, "Samples design file not specified!" } +if (params.gtf) { ch_gtf = file(params.gtf, checkIfExists: true) } else { exit 1, "GTF annotation file not specified!" } +if (params.gene_bed) { ch_gene_bed = file(params.gene_bed, checkIfExists: true) } +if (params.tss_bed) { ch_tss_bed = file(params.tss_bed, checkIfExists: true) } +if (params.blacklist) { ch_blacklist = Channel.fromPath(params.blacklist, checkIfExists: true) } else { ch_blacklist = Channel.empty() } if (params.fasta) { lastPath = params.fasta.lastIndexOf(File.separator) @@ -269,11 +269,11 @@ if (params.skip_picard_metrics) summary['Skip Picard Metrics'] = 'Yes' if (params.skip_preseq) summary['Skip Preseq'] = 'Yes' if (params.skip_plot_profile) summary['Skip plotProfile'] = 'Yes' if (params.skip_plot_fingerprint) summary['Skip plotFingerprint'] = 'Yes' -if (params.skip_ataqv) summary['Skip Ataqv'] = 'Yes' +if (params.skip_ataqv) summary['Skip Ataqv'] = 'Yes' if (params.skip_igv) summary['Skip IGV'] = 'Yes' if (params.skip_multiqc) summary['Skip MultiQC'] = 'Yes' summary['Max Resources'] = "$params.max_memory memory, $params.max_cpus cpus, $params.max_time time per job" -if (workflow.containerEngine) summary['Container'] = "$workflow.containerEngine - $workflow.container" +if (workflow.containerEngine) summary['Container'] = "$workflow.containerEngine - $workflow.container" summary['Output Dir'] = params.outdir summary['Launch Dir'] = workflow.launchDir summary['Working Dir'] = workflow.workDir @@ -901,7 +901,7 @@ process MERGED_LIB_PRESEQ { output: file "*.ccurve.txt" into ch_mlib_preseq_mqc - file "*.log" into ch_mlib_preseq_log + file "*.log" script: prefix = "${name}.mLb.mkD" @@ -940,7 +940,7 @@ process MERGED_LIB_PICARD_METRICS { output: file "*_metrics" into ch_mlib_collectmetrics_mqc - file "*.pdf" into ch_mlib_collectmetrics_pdf + file "*.pdf" script: prefix = "${name}.mLb.clN" @@ -979,8 +979,8 @@ process MERGED_LIB_BIGWIG { output: set val(name), file("*.bigWig") into ch_mlib_bigwig_plotprofile - file "*scale_factor.txt" into ch_mlib_bigwig_scale file "*igv.txt" into ch_mlib_bigwig_igv + file "*scale_factor.txt" script: prefix = "${name}.mLb.clN" @@ -1013,8 +1013,8 @@ process MERGED_LIB_PLOTPROFILE { file bed from ch_gene_bed output: - file '*.{gz,pdf}' into ch_mlib_plotprofile_results file '*.plotProfile.tab' into ch_mlib_plotprofile_mqc + file '*.{gz,pdf}' script: prefix = "${name}.mLb.clN" @@ -1052,8 +1052,8 @@ process MERGED_LIB_PLOTFINGERPRINT { set val(name), file(bam) from ch_mlib_rm_orphan_bam_plotfingerprint output: - file '*.{txt,pdf}' into ch_mlib_plotfingerprint_results file '*.raw.txt' into ch_mlib_plotfingerprint_mqc + file '*.{txt,pdf}' script: prefix = "${name}.mLb.clN" @@ -1102,13 +1102,13 @@ process MERGED_LIB_MACS2 { file mlib_frip_score_header from ch_mlib_frip_score_header output: - set val(name), file("*.{bed,xls,gappedPeak,bdg}") into ch_mlib_macs_output set val(name), file("*$PEAK_TYPE") into ch_mlib_macs_homer, ch_mlib_macs_qc, ch_mlib_macs_consensus, ch_mlib_macs_ataqv file "*igv.txt" into ch_mlib_macs_igv file "*_mqc.tsv" into ch_mlib_macs_mqc + path "*.{bed,xls,gappedPeak,bdg}" script: prefix = "${name}.mLb.clN" @@ -1183,8 +1183,8 @@ process MERGED_LIB_MACS2_QC { file mlib_peak_annotation_header from ch_mlib_peak_annotation_header output: - file "*.{txt,pdf}" into ch_mlib_peak_qc file "*.tsv" into ch_mlib_peak_qc_mqc + file "*.{txt,pdf}" script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ suffix = 'mLb.clN' @@ -1226,8 +1226,8 @@ process MERGED_LIB_CONSENSUS { file "*.bed" into ch_mlib_macs_consensus_bed file "*.saf" into ch_mlib_macs_consensus_saf file "*.boolean.txt" into ch_mlib_macs_consensus_bool - file "*.intersect.{txt,plot.pdf}" into ch_mlib_macs_consensus_intersect file "*igv.txt" into ch_mlib_macs_consensus_igv + file "*.intersect.{txt,plot.pdf}" script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ suffix = 'mLb.clN' @@ -1275,7 +1275,7 @@ process MERGED_LIB_CONSENSUS_ANNOTATE { file gtf from ch_gtf output: - file "*.annotatePeaks.txt" into ch_mlib_macs_consensus_annotate + file "*.annotatePeaks.txt" script: prefix = "consensus_peaks.mLb.clN" @@ -1348,12 +1348,12 @@ process MERGED_LIB_CONSENSUS_DESEQ2 { file mlib_deseq2_clustering_header from ch_mlib_deseq2_clustering_header output: - file "*.{RData,results.txt,pdf,log}" into ch_mlib_macs_consensus_deseq_results - file "sizeFactors" into ch_mlib_macs_consensus_deseq_factors - file "*vs*/*.{pdf,txt}" into ch_mlib_macs_consensus_deseq_comp_results - file "*vs*/*.bed" into ch_mlib_macs_consensus_deseq_comp_bed - file "*igv.txt" into ch_mlib_macs_consensus_deseq_comp_igv file "*.tsv" into ch_mlib_macs_consensus_deseq_mqc + file "*igv.txt" into ch_mlib_macs_consensus_deseq_comp_igv + file "*.{RData,results.txt,pdf,log}" + file "sizeFactors" + file "*vs*/*.{pdf,txt}" + file "*vs*/*.bed" script: prefix = "consensus_peaks.mLb.clN" @@ -1428,7 +1428,7 @@ process MERGED_LIB_ATAQV_MKARV { file json from ch_mlib_ataqv.collect() output: - file "html" into ch_mlib_ataqv_mkarv + file "html" script: """ @@ -1559,8 +1559,8 @@ process MERGED_REP_BIGWIG { output: set val(name), file("*.bigWig") into ch_mrep_bigwig - file "*scale_factor.txt" into ch_mrep_bigwig_scale file "*igv.txt" into ch_mrep_bigwig_igv + file "*scale_factor.txt" script: prefix = "${name}.mRp.clN" @@ -1599,12 +1599,12 @@ process MERGED_REP_MACS2 { file mrep_frip_score_header from ch_mrep_frip_score_header output: - set val(name), file("*.{bed,xls,gappedPeak,bdg}") into ch_mrep_macs_output set val(name), file("*$PEAK_TYPE") into ch_mrep_macs_homer, ch_mrep_macs_qc, ch_mrep_macs_consensus file "*igv.txt" into ch_mrep_macs_igv file "*_mqc.tsv" into ch_mrep_macs_mqc + path "*.{bed,xls,gappedPeak,bdg}" script: prefix = "${name}.mRp.clN" @@ -1681,7 +1681,7 @@ process MERGED_REP_MACS2_QC { output: file "*.tsv" into ch_mrep_peak_qc_mqc file "*.{txt,pdf}" - + script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ suffix = 'mRp.clN' """ @@ -1722,8 +1722,8 @@ process MERGED_REP_CONSENSUS { file "*.bed" into ch_mrep_macs_consensus_bed file "*.saf" into ch_mrep_macs_consensus_saf file "*.boolean.txt" into ch_mrep_macs_consensus_bool - file "*.intersect.{txt,plot.pdf}" into ch_mrep_macs_consensus_intersect file "*igv.txt" into ch_mrep_macs_consensus_igv + file "*.intersect.{txt,plot.pdf}" script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ suffix = 'mRp.clN' @@ -1770,7 +1770,7 @@ process MERGED_REP_CONSENSUS_ANNOTATE { file gtf from ch_gtf output: - file "*.annotatePeaks.txt" into ch_mrep_macs_consensus_annotate + file "*.annotatePeaks.txt" script: prefix = "consensus_peaks.mRp.clN" @@ -1843,12 +1843,12 @@ process MERGED_REP_CONSENSUS_DESEQ2 { file mrep_deseq2_clustering_header from ch_mrep_deseq2_clustering_header output: - file "*.{RData,results.txt,pdf,log}" into ch_mrep_macs_consensus_deseq_results - file "sizeFactors" into ch_mrep_macs_consensus_deseq_factors - file "*vs*/*.{pdf,txt}" into ch_mrep_macs_consensus_deseq_comp_results - file "*vs*/*.bed" into ch_mrep_macs_consensus_deseq_comp_bed - file "*igv.txt" into ch_mrep_macs_consensus_deseq_comp_igv file "*.tsv" into ch_mrep_macs_consensus_deseq_mqc + file "*igv.txt" into ch_mrep_macs_consensus_deseq_comp_igv + file "*.{RData,results.txt,pdf,log}" + file "sizeFactors" + file "*vs*/*.{pdf,txt}" + file "*vs*/*.bed" script: prefix = "consensus_peaks.mRp.clN" From 36e90ec07bbf658ba2a3027703b1335ae8756d6d Mon Sep 17 00:00:00 2001 From: drpatelh Date: Mon, 1 Jun 2020 23:54:49 +0100 Subject: [PATCH 180/226] Replace set with tuple --- main.nf | 70 ++++++++++++++++++++++++++++----------------------------- 1 file changed, 35 insertions(+), 35 deletions(-) diff --git a/main.nf b/main.nf index 304ac497..6b41a14a 100644 --- a/main.nf +++ b/main.nf @@ -501,7 +501,7 @@ process FASTQC { !params.skip_fastqc input: - set val(name), file(reads) from ch_raw_reads_fastqc + tuple val(name), path(reads) from ch_raw_reads_fastqc output: file "*.{zip,html}" into ch_fastqc_reports_mqc @@ -551,10 +551,10 @@ if (params.skip_trimming) { } input: - set val(name), file(reads) from ch_raw_reads_trimgalore + tuple val(name), path(reads) from ch_raw_reads_trimgalore output: - set val(name), file("*.fq.gz") into ch_trimmed_reads + tuple val(name), path("*.fq.gz") into ch_trimmed_reads file "*.txt" into ch_trimgalore_results_mqc file "*.{zip,html}" into ch_trimgalore_fastqc_reports_mqc @@ -609,11 +609,11 @@ process BWA_MEM { label 'process_high' input: - set val(name), file(reads) from ch_trimmed_reads + tuple val(name), path(reads) from ch_trimmed_reads file index from ch_bwa_index.collect() output: - set val(name), file("*.bam") into ch_bwa_bam + tuple val(name), path("*.bam") into ch_bwa_bam script: prefix = "${name}.Lb" @@ -649,10 +649,10 @@ process SORT_BAM { } input: - set val(name), file(bam) from ch_bwa_bam + tuple val(name), path(bam) from ch_bwa_bam output: - set val(name), file("*.sorted.{bam,bam.bai}") into ch_sort_bam_merge + tuple val(name), path("*.sorted.{bam,bam.bai}") into ch_sort_bam_merge file "*.{flagstat,idxstats,stats}" into ch_sort_bam_flagstat_mqc script: @@ -696,10 +696,10 @@ process MERGED_LIB_BAM { } input: - set val(name), file(bams) from ch_sort_bam_merge + tuple val(name), path(bams) from ch_sort_bam_merge output: - set val(name), file("*${prefix}.sorted.{bam,bam.bai}") into ch_mlib_bam_filter, + tuple val(name), path("*${prefix}.sorted.{bam,bam.bai}") into ch_mlib_bam_filter, ch_mlib_bam_preseq, ch_mlib_bam_ataqv file "*.{flagstat,idxstats,stats}" into ch_mlib_bam_stats_mqc @@ -776,13 +776,13 @@ process MERGED_LIB_BAM_FILTER { } input: - set val(name), file(bam) from ch_mlib_bam_filter + tuple val(name), path(bam) from ch_mlib_bam_filter file bed from ch_genome_filter_regions.collect() file bamtools_filter_config from ch_bamtools_filter_config output: - set val(name), file("*.{bam,bam.bai}") into ch_mlib_filter_bam - set val(name), file("*.flagstat") into ch_mlib_filter_bam_flagstat + tuple val(name), path("*.{bam,bam.bai}") into ch_mlib_filter_bam + tuple val(name), path("*.flagstat") into ch_mlib_filter_bam_flagstat file "*.{idxstats,stats}" into ch_mlib_filter_bam_stats_mqc script: @@ -847,17 +847,17 @@ if (params.single_end) { } input: - set val(name), file(bam) from ch_mlib_filter_bam + tuple val(name), path(bam) from ch_mlib_filter_bam output: - set val(name), file("*.sorted.{bam,bam.bai}") into ch_mlib_rm_orphan_bam_metrics, + tuple val(name), path("*.sorted.{bam,bam.bai}") into ch_mlib_rm_orphan_bam_metrics, ch_mlib_rm_orphan_bam_bigwig, ch_mlib_rm_orphan_bam_macs, ch_mlib_rm_orphan_bam_plotfingerprint, ch_mlib_rm_orphan_bam_mrep - set val(name), file("${prefix}.bam") into ch_mlib_name_bam_mlib_counts, + tuple val(name), path("${prefix}.bam") into ch_mlib_name_bam_mlib_counts, ch_mlib_name_bam_mrep_counts - set val(name), file("*.flagstat") into ch_mlib_rm_orphan_flagstat_bigwig, + tuple val(name), path("*.flagstat") into ch_mlib_rm_orphan_flagstat_bigwig, ch_mlib_rm_orphan_flagstat_macs, ch_mlib_rm_orphan_flagstat_mqc file "*.{idxstats,stats}" into ch_mlib_rm_orphan_stats_mqc @@ -897,7 +897,7 @@ process MERGED_LIB_PRESEQ { !params.skip_preseq input: - set val(name), file(bam) from ch_mlib_bam_preseq + tuple val(name), path(bam) from ch_mlib_bam_preseq output: file "*.ccurve.txt" into ch_mlib_preseq_mqc @@ -935,7 +935,7 @@ process MERGED_LIB_PICARD_METRICS { !params.skip_picard_metrics input: - set val(name), file(bam) from ch_mlib_rm_orphan_bam_metrics + tuple val(name), path(bam) from ch_mlib_rm_orphan_bam_metrics file fasta from ch_fasta output: @@ -974,11 +974,11 @@ process MERGED_LIB_BIGWIG { } input: - set val(name), file(bam), file(flagstat) from ch_mlib_rm_orphan_bam_bigwig.join(ch_mlib_rm_orphan_flagstat_bigwig, by: [0]) + tuple val(name), path(bam), path(flagstat) from ch_mlib_rm_orphan_bam_bigwig.join(ch_mlib_rm_orphan_flagstat_bigwig, by: [0]) file sizes from ch_genome_sizes_mlib_bigwig.collect() output: - set val(name), file("*.bigWig") into ch_mlib_bigwig_plotprofile + tuple val(name), path("*.bigWig") into ch_mlib_bigwig_plotprofile file "*igv.txt" into ch_mlib_bigwig_igv file "*scale_factor.txt" @@ -1009,7 +1009,7 @@ process MERGED_LIB_PLOTPROFILE { !params.skip_plot_profile input: - set val(name), file(bigwig) from ch_mlib_bigwig_plotprofile + tuple val(name), path(bigwig) from ch_mlib_bigwig_plotprofile file bed from ch_gene_bed output: @@ -1049,7 +1049,7 @@ process MERGED_LIB_PLOTFINGERPRINT { !params.skip_plot_fingerprint input: - set val(name), file(bam) from ch_mlib_rm_orphan_bam_plotfingerprint + tuple val(name), path(bam) from ch_mlib_rm_orphan_bam_plotfingerprint output: file '*.raw.txt' into ch_mlib_plotfingerprint_mqc @@ -1097,12 +1097,12 @@ process MERGED_LIB_MACS2 { params.macs_gsize input: - set val(name), file(bam), file(flagstat) from ch_mlib_rm_orphan_bam_macs.join(ch_mlib_rm_orphan_flagstat_macs, by: [0]) + tuple val(name), path(bam), path(flagstat) from ch_mlib_rm_orphan_bam_macs.join(ch_mlib_rm_orphan_flagstat_macs, by: [0]) file mlib_peak_count_header from ch_mlib_peak_count_header file mlib_frip_score_header from ch_mlib_frip_score_header output: - set val(name), file("*$PEAK_TYPE") into ch_mlib_macs_homer, + tuple val(name), path("*$PEAK_TYPE") into ch_mlib_macs_homer, ch_mlib_macs_qc, ch_mlib_macs_consensus, ch_mlib_macs_ataqv @@ -1147,7 +1147,7 @@ process MERGED_LIB_MACS2_ANNOTATE { params.macs_gsize input: - set val(name), file(peak) from ch_mlib_macs_homer + tuple val(name), path(peak) from ch_mlib_macs_homer file fasta from ch_fasta file gtf from ch_gtf @@ -1388,7 +1388,7 @@ process MERGED_LIB_ATAQV { !params.skip_ataqv input: - set val(name), file(bam), file(peak) from ch_mlib_bam_ataqv.join(ch_mlib_macs_ataqv, by: [0]) + tuple val(name), path(bam), path(peak) from ch_mlib_bam_ataqv.join(ch_mlib_macs_ataqv, by: [0]) file autosomes from ch_genome_autosomes.collect() file tss_bed from ch_tss_bed @@ -1470,12 +1470,12 @@ process MERGED_REP_BAM { } input: - set val(name), file(bams) from ch_mlib_rm_orphan_bam_mrep + tuple val(name), path(bams) from ch_mlib_rm_orphan_bam_mrep output: - set val(name), file("*${prefix}.sorted.{bam,bam.bai}") into ch_mrep_bam_bigwig, + tuple val(name), path("*${prefix}.sorted.{bam,bam.bai}") into ch_mrep_bam_bigwig, ch_mrep_bam_macs - set val(name), file("*.flagstat") into ch_mrep_bam_flagstat_bigwig, + tuple val(name), path("*.flagstat") into ch_mrep_bam_flagstat_bigwig, ch_mrep_bam_flagstat_macs, ch_mrep_bam_flagstat_mqc file "*.{idxstats,stats}" into ch_mrep_bam_stats_mqc @@ -1554,11 +1554,11 @@ process MERGED_REP_BIGWIG { !params.skip_merge_replicates && replicatesExist input: - set val(name), file(bam), file(flagstat) from ch_mrep_bam_bigwig.join(ch_mrep_bam_flagstat_bigwig, by: [0]) + tuple val(name), path(bam), path(flagstat) from ch_mrep_bam_bigwig.join(ch_mrep_bam_flagstat_bigwig, by: [0]) file sizes from ch_genome_sizes_mrep_bigwig.collect() output: - set val(name), file("*.bigWig") into ch_mrep_bigwig + tuple val(name), path("*.bigWig") into ch_mrep_bigwig file "*igv.txt" into ch_mrep_bigwig_igv file "*scale_factor.txt" @@ -1594,12 +1594,12 @@ process MERGED_REP_MACS2 { !params.skip_merge_replicates && replicatesExist && params.macs_gsize input: - set val(name), file(bam), file(flagstat) from ch_mrep_bam_macs.join(ch_mrep_bam_flagstat_macs, by: [0]) + tuple val(name), path(bam), path(flagstat) from ch_mrep_bam_macs.join(ch_mrep_bam_flagstat_macs, by: [0]) file mrep_peak_count_header from ch_mrep_peak_count_header file mrep_frip_score_header from ch_mrep_frip_score_header output: - set val(name), file("*$PEAK_TYPE") into ch_mrep_macs_homer, + tuple val(name), path("*$PEAK_TYPE") into ch_mrep_macs_homer, ch_mrep_macs_qc, ch_mrep_macs_consensus file "*igv.txt" into ch_mrep_macs_igv @@ -1643,7 +1643,7 @@ process MERGED_REP_MACS2_ANNOTATE { !params.skip_merge_replicates && replicatesExist && params.macs_gsize input: - set val(name), file(peak) from ch_mrep_macs_homer + tuple val(name), path(peak) from ch_mrep_macs_homer file fasta from ch_fasta file gtf from ch_gtf @@ -1902,7 +1902,7 @@ process IGV { file rdifferential_peaks from ch_mrep_macs_consensus_deseq_comp_igv.collect().ifEmpty([]) output: - file "*.{txt,xml}" into ch_igv_session + file "*.{txt,xml}" script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ """ From a84a5117667ac4e5eae56d58f9d8b86226935b8b Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 2 Jun 2020 10:00:21 +0100 Subject: [PATCH 181/226] Unify syntax --- main.nf | 578 ++++++++++++++++++++++++++++---------------------------- 1 file changed, 289 insertions(+), 289 deletions(-) diff --git a/main.nf b/main.nf index 6b41a14a..82fb4ee4 100644 --- a/main.nf +++ b/main.nf @@ -135,7 +135,7 @@ params.blacklist = params.genome ? params.genomes[ params.genome ].blacklist ?: params.anno_readme = params.genome ? params.genomes[ params.genome ].readme ?: false : false // Global variables -def PEAK_TYPE = params.narrow_peak ? "narrowPeak" : "broadPeak" +def PEAK_TYPE = params.narrow_peak ? 'narrowPeak' : 'broadPeak' //////////////////////////////////////////////////// /* -- CONFIG FILES -- */ @@ -171,8 +171,8 @@ ch_mrep_deseq2_clustering_header = file("$baseDir/assets/multiqc/mrep_deseq2_clu /* -- VALIDATE INPUTS -- */ //////////////////////////////////////////////////// -if (params.input) { ch_input = file(params.input, checkIfExists: true) } else { exit 1, "Samples design file not specified!" } -if (params.gtf) { ch_gtf = file(params.gtf, checkIfExists: true) } else { exit 1, "GTF annotation file not specified!" } +if (params.input) { ch_input = file(params.input, checkIfExists: true) } else { exit 1, 'Samples design file not specified!' } +if (params.gtf) { ch_gtf = file(params.gtf, checkIfExists: true) } else { exit 1, 'GTF annotation file not specified!' } if (params.gene_bed) { ch_gene_bed = file(params.gene_bed, checkIfExists: true) } if (params.tss_bed) { ch_tss_bed = file(params.tss_bed, checkIfExists: true) } if (params.blacklist) { ch_blacklist = Channel.fromPath(params.blacklist, checkIfExists: true) } else { ch_blacklist = Channel.empty() } @@ -182,7 +182,7 @@ if (params.fasta) { bwa_base = params.fasta.substring(lastPath+1) ch_fasta = file(params.fasta, checkIfExists: true) } else { - exit 1, "Fasta file not specified!" + exit 1, 'Fasta file not specified!' } if (params.bwa_index) { @@ -206,12 +206,12 @@ if (params.anno_readme && file(params.anno_readme).exists()) { if (workflow.profile.contains('awsbatch')) { // AWSBatch sanity checking - if (!params.awsqueue || !params.awsregion) exit 1, "Specify correct --awsqueue and --awsregion parameters on AWSBatch!" + if (!params.awsqueue || !params.awsregion) exit 1, 'Specify correct --awsqueue and --awsregion parameters on AWSBatch!' // Check outdir paths to be S3 buckets if running on AWSBatch // related: https://github.com/nextflow-io/nextflow/issues/813 - if (!params.outdir.startsWith('s3:')) exit 1, "Outdir not on S3 - specify S3 Bucket to run on AWSBatch!" + if (!params.outdir.startsWith('s3:')) exit 1, 'Outdir not on S3 - specify S3 Bucket to run on AWSBatch!' // Prevent trace files to be stored on S3 since S3 does not support rolling files. - if (params.tracedir.startsWith('s3:')) exit 1, "Specify a local tracedir or run without trace! S3 cannot be used for tracefiles." + if (params.tracedir.startsWith('s3:')) exit 1, 'Specify a local tracedir or run without trace! S3 cannot be used for tracefiles.' } /////////////////////////////////////////////////////////////////////////////// @@ -247,7 +247,7 @@ if (params.skip_trimming) { summary['Trim R2'] = "$params.clip_r2 bp" summary["Trim 3' R1"] = "$params.three_prime_clip_r1 bp" summary["Trim 3' R2"] = "$params.three_prime_clip_r2 bp" - summary["NextSeq Trim"] = "$params.trim_nextseq bp" + summary['NextSeq Trim'] = "$params.trim_nextseq bp" } if (params.seq_center) summary['Sequencing Center'] = params.seq_center if (params.single_end) summary['Fragment Size'] = "$params.fragment_size bp" @@ -293,7 +293,7 @@ if (params.email || params.email_on_fail) { summary['E-mail on failure'] = params.email_on_fail summary['MultiQC Max Size'] = params.max_multiqc_email_size } -log.info summary.collect { k,v -> "${k.padRight(21)}: $v" }.join("\n") +log.info summary.collect { k,v -> "${k.padRight(21)}: $v" }.join('\n') log.info "-\033[2m--------------------------------------------------\033[0m-" // Check the hostnames against configured profiles @@ -325,10 +325,10 @@ process CHECK_DESIGN { publishDir "${params.outdir}/pipeline_info", mode: params.publish_dir_mode input: - file design from ch_input + path design from ch_input output: - file "*.csv" into ch_design_reads_csv + path '*.csv' into ch_design_reads_csv script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ """ @@ -391,10 +391,10 @@ if (!params.bwa_index) { saveAs: { params.save_reference ? it : null }, mode: params.publish_dir_mode input: - file fasta from ch_fasta + path fasta from ch_fasta output: - file "BWAIndex" into ch_bwa_index + path 'BWAIndex' into ch_bwa_index script: """ @@ -414,10 +414,10 @@ if (!params.gene_bed) { publishDir "${params.outdir}/genome", mode: params.publish_dir_mode input: - file gtf from ch_gtf + path gtf from ch_gtf output: - file "*.bed" into ch_gene_bed + path '*.bed' into ch_gene_bed script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ """ @@ -435,10 +435,10 @@ if (!params.tss_bed) { publishDir "${params.outdir}/genome", mode: params.publish_dir_mode input: - file bed from ch_gene_bed + path bed from ch_gene_bed output: - file "*.bed" into ch_tss_bed + path '*.bed' into ch_tss_bed script: """ @@ -455,21 +455,21 @@ process MAKE_GENOME_FILTER { publishDir "${params.outdir}/genome", mode: params.publish_dir_mode input: - file fasta from ch_fasta - file blacklist from ch_blacklist.ifEmpty([]) + path fasta from ch_fasta + path blacklist from ch_blacklist.ifEmpty([]) output: - file "$fasta" // FASTA FILE FOR IGV - file "*.fai" // FAI INDEX FOR REFERENCE GENOME - file "*.bed" into ch_genome_filter_regions // BED FILE WITHOUT BLACKLIST REGIONS & MITOCHONDRIAL CONTIG FOR FILTERING - file "*.txt" into ch_genome_autosomes // TEXT FILE CONTAINING LISTING OF AUTOSOMAL CHROMOSOMES FOR ATAQV - file "*.sizes" into ch_genome_sizes_mlib_bigwig, // CHROMOSOME SIZES FILE FOR BEDTOOLS + path "$fasta" // FASTA FILE FOR IGV + path '*.fai' // FAI INDEX FOR REFERENCE GENOME + path '*.bed' into ch_genome_filter_regions // BED FILE WITHOUT BLACKLIST REGIONS & MITOCHONDRIAL CONTIG FOR FILTERING + path '*.txt' into ch_genome_autosomes // TEXT FILE CONTAINING LISTING OF AUTOSOMAL CHROMOSOMES FOR ATAQV + path '*.sizes' into ch_genome_sizes_mlib_bigwig, // CHROMOSOME SIZES FILE FOR BEDTOOLS ch_genome_sizes_mrep_bigwig script: blacklist_filter = params.blacklist ? "sortBed -i $blacklist -g ${fasta}.sizes | complementBed -i stdin -g ${fasta}.sizes" : "awk '{print \$1, '0' , \$2}' OFS='\t' ${fasta}.sizes" - name_filter = params.mito_name ? "| awk '\$1 !~ /${params.mito_name}/ {print \$0}'": "" - mito_filter = params.keep_mito ? "" : name_filter + name_filter = params.mito_name ? "| awk '\$1 !~ /${params.mito_name}/ {print \$0}'": '' + mito_filter = params.keep_mito ? '' : name_filter """ samtools faidx $fasta get_autosomes.py ${fasta}.fai ${fasta}.autosomes.txt @@ -494,7 +494,7 @@ process FASTQC { label 'process_medium' publishDir "${params.outdir}/fastqc", mode: params.publish_dir_mode, saveAs: { filename -> - filename.endsWith(".zip") ? "zips/$filename" : "$filename" + filename.endsWith('.zip') ? "zips/$filename" : filename } when: @@ -504,7 +504,7 @@ process FASTQC { tuple val(name), path(reads) from ch_raw_reads_fastqc output: - file "*.{zip,html}" into ch_fastqc_reports_mqc + path '*.{zip,html}' into ch_fastqc_reports_mqc script: // Added soft-links to original fastqs for consistent naming in MultiQC @@ -544,9 +544,9 @@ if (params.skip_trimming) { label 'process_high' publishDir "${params.outdir}/trim_galore", mode: params.publish_dir_mode, saveAs: { filename -> - if (filename.endsWith(".html")) "fastqc/$filename" - else if (filename.endsWith(".zip")) "fastqc/zips/$filename" - else if (filename.endsWith("trimming_report.txt")) "logs/$filename" + if (filename.endsWith('.html')) "fastqc/$filename" + else if (filename.endsWith('.zip')) "fastqc/zips/$filename" + else if (filename.endsWith('trimming_report.txt')) "logs/$filename" else params.save_trimmed ? filename : null } @@ -554,9 +554,9 @@ if (params.skip_trimming) { tuple val(name), path(reads) from ch_raw_reads_trimgalore output: - tuple val(name), path("*.fq.gz") into ch_trimmed_reads - file "*.txt" into ch_trimgalore_results_mqc - file "*.{zip,html}" into ch_trimgalore_fastqc_reports_mqc + tuple val(name), path('*.fq.gz') into ch_trimmed_reads + path '*.txt' into ch_trimgalore_results_mqc + path '*.{zip,html}' into ch_trimgalore_fastqc_reports_mqc script: // Calculate number of --cores for TrimGalore based on value of task.cpus @@ -610,10 +610,10 @@ process BWA_MEM { input: tuple val(name), path(reads) from ch_trimmed_reads - file index from ch_bwa_index.collect() + path index from ch_bwa_index.collect() output: - tuple val(name), path("*.bam") into ch_bwa_bam + tuple val(name), path('*.bam') into ch_bwa_bam script: prefix = "${name}.Lb" @@ -641,9 +641,9 @@ process SORT_BAM { if (params.save_align_intermeds) { publishDir path: "${params.outdir}/bwa/library", mode: params.publish_dir_mode, saveAs: { filename -> - if (filename.endsWith(".flagstat")) "samtools_stats/$filename" - else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" - else if (filename.endsWith(".stats")) "samtools_stats/$filename" + if (filename.endsWith('.flagstat')) "samtools_stats/$filename" + else if (filename.endsWith('.idxstats')) "samtools_stats/$filename" + else if (filename.endsWith('.stats')) "samtools_stats/$filename" else filename } } @@ -652,8 +652,8 @@ process SORT_BAM { tuple val(name), path(bam) from ch_bwa_bam output: - tuple val(name), path("*.sorted.{bam,bam.bai}") into ch_sort_bam_merge - file "*.{flagstat,idxstats,stats}" into ch_sort_bam_flagstat_mqc + tuple val(name), path('*.sorted.{bam,bam.bai}') into ch_sort_bam_merge + path '*.{flagstat,idxstats,stats}' into ch_sort_bam_flagstat_mqc script: prefix = "${name}.Lb" @@ -688,10 +688,10 @@ process MERGED_LIB_BAM { label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary", mode: params.publish_dir_mode, saveAs: { filename -> - if (filename.endsWith(".flagstat")) "samtools_stats/$filename" - else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" - else if (filename.endsWith(".stats")) "samtools_stats/$filename" - else if (filename.endsWith(".metrics.txt")) "picard_metrics/$filename" + if (filename.endsWith('.flagstat')) "samtools_stats/$filename" + else if (filename.endsWith('.idxstats')) "samtools_stats/$filename" + else if (filename.endsWith('.stats')) "samtools_stats/$filename" + else if (filename.endsWith('.metrics.txt')) "picard_metrics/$filename" else params.save_align_intermeds ? filename : null } @@ -700,17 +700,17 @@ process MERGED_LIB_BAM { output: tuple val(name), path("*${prefix}.sorted.{bam,bam.bai}") into ch_mlib_bam_filter, - ch_mlib_bam_preseq, - ch_mlib_bam_ataqv - file "*.{flagstat,idxstats,stats}" into ch_mlib_bam_stats_mqc - file "*.txt" into ch_mlib_bam_metrics_mqc + ch_mlib_bam_preseq, + ch_mlib_bam_ataqv + path '*.{flagstat,idxstats,stats}' into ch_mlib_bam_stats_mqc + path '*.txt' into ch_mlib_bam_metrics_mqc script: prefix = "${name}.mLb.mkD" bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() def avail_mem = 3 if (!task.memory) { - log.info "[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." + log.info '[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this.' } else { avail_mem = task.memory.toGiga() } @@ -766,32 +766,32 @@ process MERGED_LIB_BAM_FILTER { publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: params.publish_dir_mode, saveAs: { filename -> if (params.single_end || params.save_align_intermeds) { - if (filename.endsWith(".flagstat")) "samtools_stats/$filename" - else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" - else if (filename.endsWith(".stats")) "samtools_stats/$filename" - else if (filename.endsWith(".sorted.bam")) filename - else if (filename.endsWith(".sorted.bam.bai")) filename + if (filename.endsWith('.flagstat')) "samtools_stats/$filename" + else if (filename.endsWith('.idxstats')) "samtools_stats/$filename" + else if (filename.endsWith('.stats')) "samtools_stats/$filename" + else if (filename.endsWith('.sorted.bam')) filename + else if (filename.endsWith('.sorted.bam.bai')) filename else null } } input: tuple val(name), path(bam) from ch_mlib_bam_filter - file bed from ch_genome_filter_regions.collect() - file bamtools_filter_config from ch_bamtools_filter_config + path bed from ch_genome_filter_regions.collect() + path bamtools_filter_config from ch_bamtools_filter_config output: - tuple val(name), path("*.{bam,bam.bai}") into ch_mlib_filter_bam - tuple val(name), path("*.flagstat") into ch_mlib_filter_bam_flagstat - file "*.{idxstats,stats}" into ch_mlib_filter_bam_stats_mqc + tuple val(name), path('*.{bam,bam.bai}') into ch_mlib_filter_bam + tuple val(name), path('*.flagstat') into ch_mlib_filter_bam_flagstat + path '*.{idxstats,stats}' into ch_mlib_filter_bam_stats_mqc script: prefix = params.single_end ? "${name}.mLb.clN" : "${name}.mLb.flT" - filter_params = params.single_end ? "-F 0x004" : "-F 0x004 -F 0x0008 -f 0x001" - dup_params = params.keep_dups ? "" : "-F 0x0400" - multimap_params = params.keep_multi_map ? "" : "-q 1" - blacklist_params = params.blacklist ? "-L $bed" : "" - name_sort_bam = params.single_end ? "" : "samtools sort -n -@ $task.cpus -o ${prefix}.bam -T $prefix ${prefix}.sorted.bam" + filter_params = params.single_end ? '-F 0x004' : '-F 0x004 -F 0x0008 -f 0x001' + dup_params = params.keep_dups ? '' : '-F 0x0400' + multimap_params = params.keep_multi_map ? '' : '-q 1' + blacklist_params = params.blacklist ? "-L $bed" : '' + name_sort_bam = params.single_end ? '' : "samtools sort -n -@ $task.cpus -o ${prefix}.bam -T $prefix ${prefix}.sorted.bam" """ samtools view \\ $filter_params \\ @@ -838,11 +838,11 @@ if (params.single_end) { label 'process_medium' publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: params.publish_dir_mode, saveAs: { filename -> - if (filename.endsWith(".flagstat")) "samtools_stats/$filename" - else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" - else if (filename.endsWith(".stats")) "samtools_stats/$filename" - else if (filename.endsWith(".sorted.bam")) filename - else if (filename.endsWith(".sorted.bam.bai")) filename + if (filename.endsWith('.flagstat')) "samtools_stats/$filename" + else if (filename.endsWith('.idxstats')) "samtools_stats/$filename" + else if (filename.endsWith('.stats')) "samtools_stats/$filename" + else if (filename.endsWith('.sorted.bam')) filename + else if (filename.endsWith('.sorted.bam.bai')) filename else null } @@ -850,17 +850,17 @@ if (params.single_end) { tuple val(name), path(bam) from ch_mlib_filter_bam output: - tuple val(name), path("*.sorted.{bam,bam.bai}") into ch_mlib_rm_orphan_bam_metrics, - ch_mlib_rm_orphan_bam_bigwig, - ch_mlib_rm_orphan_bam_macs, - ch_mlib_rm_orphan_bam_plotfingerprint, - ch_mlib_rm_orphan_bam_mrep + tuple val(name), path('*.sorted.{bam,bam.bai}') into ch_mlib_rm_orphan_bam_metrics, + ch_mlib_rm_orphan_bam_bigwig, + ch_mlib_rm_orphan_bam_macs, + ch_mlib_rm_orphan_bam_plotfingerprint, + ch_mlib_rm_orphan_bam_mrep tuple val(name), path("${prefix}.bam") into ch_mlib_name_bam_mlib_counts, - ch_mlib_name_bam_mrep_counts - tuple val(name), path("*.flagstat") into ch_mlib_rm_orphan_flagstat_bigwig, - ch_mlib_rm_orphan_flagstat_macs, - ch_mlib_rm_orphan_flagstat_mqc - file "*.{idxstats,stats}" into ch_mlib_rm_orphan_stats_mqc + ch_mlib_name_bam_mrep_counts + tuple val(name), path('*.flagstat') into ch_mlib_rm_orphan_flagstat_bigwig, + ch_mlib_rm_orphan_flagstat_macs, + ch_mlib_rm_orphan_flagstat_mqc + path '*.{idxstats,stats}' into ch_mlib_rm_orphan_stats_mqc script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ prefix = "${name}.mLb.clN" @@ -900,12 +900,12 @@ process MERGED_LIB_PRESEQ { tuple val(name), path(bam) from ch_mlib_bam_preseq output: - file "*.ccurve.txt" into ch_mlib_preseq_mqc - file "*.log" + path '*.ccurve.txt' into ch_mlib_preseq_mqc + path '*.log' script: prefix = "${name}.mLb.mkD" - pe = params.single_end ? "" : "-pe" + pe = params.single_end ? '' : '-pe' """ preseq lc_extrap \\ -output ${prefix}.ccurve.txt \\ @@ -926,8 +926,8 @@ process MERGED_LIB_PICARD_METRICS { label 'process_medium' publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: params.publish_dir_mode, saveAs: { filename -> - if (filename.endsWith("_metrics")) "picard_metrics/$filename" - else if (filename.endsWith(".pdf")) "picard_metrics/pdf/$filename" + if (filename.endsWith('_metrics')) "picard_metrics/$filename" + else if (filename.endsWith('.pdf')) "picard_metrics/pdf/$filename" else null } @@ -936,17 +936,17 @@ process MERGED_LIB_PICARD_METRICS { input: tuple val(name), path(bam) from ch_mlib_rm_orphan_bam_metrics - file fasta from ch_fasta + path fasta from ch_fasta output: - file "*_metrics" into ch_mlib_collectmetrics_mqc - file "*.pdf" + path '*_metrics' into ch_mlib_collectmetrics_mqc + path '*.pdf' script: prefix = "${name}.mLb.clN" def avail_mem = 3 if (!task.memory) { - log.info "[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." + log.info '[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this.' } else { avail_mem = task.memory.toGiga() } @@ -968,23 +968,23 @@ process MERGED_LIB_BIGWIG { label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/bigwig", mode: params.publish_dir_mode, saveAs: { filename -> - if (filename.endsWith("scale_factor.txt")) "scale/$filename" - else if (filename.endsWith(".bigWig")) "$filename" + if (filename.endsWith('scale_factor.txt')) "scale/$filename" + else if (filename.endsWith('.bigWig')) filename else null } input: tuple val(name), path(bam), path(flagstat) from ch_mlib_rm_orphan_bam_bigwig.join(ch_mlib_rm_orphan_flagstat_bigwig, by: [0]) - file sizes from ch_genome_sizes_mlib_bigwig.collect() + path sizes from ch_genome_sizes_mlib_bigwig.collect() output: - tuple val(name), path("*.bigWig") into ch_mlib_bigwig_plotprofile - file "*igv.txt" into ch_mlib_bigwig_igv - file "*scale_factor.txt" + tuple val(name), path('*.bigWig') into ch_mlib_bigwig_plotprofile + path '*igv.txt' into ch_mlib_bigwig_igv + path '*scale_factor.txt' script: prefix = "${name}.mLb.clN" - pe_fragment = params.single_end ? "" : "-pc" + pe_fragment = params.single_end ? '' : '-pc' extend = (params.single_end && params.fragment_size > 0) ? "-fs ${params.fragment_size}" : '' """ SCALE_FACTOR=\$(grep 'mapped (' $flagstat | awk '{print 1000000/\$1}') @@ -1010,11 +1010,11 @@ process MERGED_LIB_PLOTPROFILE { input: tuple val(name), path(bigwig) from ch_mlib_bigwig_plotprofile - file bed from ch_gene_bed + path bed from ch_gene_bed output: - file '*.plotProfile.tab' into ch_mlib_plotprofile_mqc - file '*.{gz,pdf}' + path '*.plotProfile.tab' into ch_mlib_plotprofile_mqc + path '*.{gz,pdf}' script: prefix = "${name}.mLb.clN" @@ -1052,8 +1052,8 @@ process MERGED_LIB_PLOTFINGERPRINT { tuple val(name), path(bam) from ch_mlib_rm_orphan_bam_plotfingerprint output: - file '*.raw.txt' into ch_mlib_plotfingerprint_mqc - file '*.{txt,pdf}' + path '*.raw.txt' into ch_mlib_plotfingerprint_mqc + path '*.{txt,pdf}' script: prefix = "${name}.mLb.clN" @@ -1088,8 +1088,8 @@ process MERGED_LIB_MACS2 { label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}", mode: params.publish_dir_mode, saveAs: { filename -> - if (filename.endsWith(".tsv")) "qc/$filename" - else if (filename.endsWith(".igv.txt")) null + if (filename.endsWith('.tsv')) "qc/$filename" + else if (filename.endsWith('.igv.txt')) null else filename } @@ -1098,23 +1098,23 @@ process MERGED_LIB_MACS2 { input: tuple val(name), path(bam), path(flagstat) from ch_mlib_rm_orphan_bam_macs.join(ch_mlib_rm_orphan_flagstat_macs, by: [0]) - file mlib_peak_count_header from ch_mlib_peak_count_header - file mlib_frip_score_header from ch_mlib_frip_score_header + path mlib_peak_count_header from ch_mlib_peak_count_header + path mlib_frip_score_header from ch_mlib_frip_score_header output: tuple val(name), path("*$PEAK_TYPE") into ch_mlib_macs_homer, - ch_mlib_macs_qc, - ch_mlib_macs_consensus, - ch_mlib_macs_ataqv - file "*igv.txt" into ch_mlib_macs_igv - file "*_mqc.tsv" into ch_mlib_macs_mqc - path "*.{bed,xls,gappedPeak,bdg}" + ch_mlib_macs_qc, + ch_mlib_macs_consensus, + ch_mlib_macs_ataqv + path '*igv.txt' into ch_mlib_macs_igv + path '*_mqc.tsv' into ch_mlib_macs_mqc + path '*.{bed,xls,gappedPeak,bdg}' script: prefix = "${name}.mLb.clN" broad = params.narrow_peak ? '' : "--broad --broad-cutoff ${params.broad_cutoff}" - format = params.single_end ? "BAM" : "BAMPE" - pileup = params.save_macs_pileup ? "-B --SPMR" : "" + format = params.single_end ? 'BAM' : 'BAMPE' + pileup = params.save_macs_pileup ? '-B --SPMR' : '' """ macs2 callpeak \\ -t ${bam[0]} \\ @@ -1140,7 +1140,7 @@ process MERGED_LIB_MACS2 { */ process MERGED_LIB_MACS2_ANNOTATE { tag "$name" - label "process_medium" + label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}", mode: params.publish_dir_mode when: @@ -1148,11 +1148,11 @@ process MERGED_LIB_MACS2_ANNOTATE { input: tuple val(name), path(peak) from ch_mlib_macs_homer - file fasta from ch_fasta - file gtf from ch_gtf + path fasta from ch_fasta + path gtf from ch_gtf output: - file "*.txt" into ch_mlib_macs_annotate + path '*.txt' into ch_mlib_macs_annotate script: prefix = "${name}.mLb.clN" @@ -1171,20 +1171,20 @@ process MERGED_LIB_MACS2_ANNOTATE { * STEP 6.3: Aggregated QC plots for peaks, FRiP and peak-to-gene annotation */ process MERGED_LIB_MACS2_QC { - label "process_medium" + label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/qc", mode: params.publish_dir_mode when: params.macs_gsize && !params.skip_peak_qc input: - file peaks from ch_mlib_macs_qc.collect{ it[1] } - file annos from ch_mlib_macs_annotate.collect() - file mlib_peak_annotation_header from ch_mlib_peak_annotation_header + path peaks from ch_mlib_macs_qc.collect{ it[1] } + path annos from ch_mlib_macs_annotate.collect() + path mlib_peak_annotation_header from ch_mlib_peak_annotation_header output: - file "*.tsv" into ch_mlib_peak_qc_mqc - file "*.{txt,pdf}" + path '*.tsv' into ch_mlib_peak_qc_mqc + path '*.{txt,pdf}' script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ suffix = 'mLb.clN' @@ -1212,7 +1212,7 @@ process MERGED_LIB_CONSENSUS { label 'process_long' publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: params.publish_dir_mode, saveAs: { filename -> - if (filename.endsWith(".igv.txt")) null + if (filename.endsWith('.igv.txt')) null else filename } @@ -1220,21 +1220,21 @@ process MERGED_LIB_CONSENSUS { params.macs_gsize && (replicatesExist || multipleGroups) && !params.skip_consensus_peaks input: - file peaks from ch_mlib_macs_consensus.collect{ it[1] } + path peaks from ch_mlib_macs_consensus.collect{ it[1] } output: - file "*.bed" into ch_mlib_macs_consensus_bed - file "*.saf" into ch_mlib_macs_consensus_saf - file "*.boolean.txt" into ch_mlib_macs_consensus_bool - file "*igv.txt" into ch_mlib_macs_consensus_igv - file "*.intersect.{txt,plot.pdf}" + path '*.bed' into ch_mlib_macs_consensus_bed + path '*.saf' into ch_mlib_macs_consensus_saf + path '*.boolean.txt' into ch_mlib_macs_consensus_bool + path '*igv.txt' into ch_mlib_macs_consensus_igv + path '*.intersect.{txt,plot.pdf}' script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ suffix = 'mLb.clN' prefix = "consensus_peaks.${suffix}" mergecols = params.narrow_peak ? (2..10).join(',') : (2..9).join(',') - collapsecols = params.narrow_peak ? (["collapse"]*9).join(',') : (["collapse"]*8).join(',') - expandparam = params.narrow_peak ? "--is_narrow_peak" : "" + collapsecols = params.narrow_peak ? (['collapse']*9).join(',') : (['collapse']*8).join(',') + expandparam = params.narrow_peak ? '--is_narrow_peak' : '' """ sort -T '.' -k1,1 -k2,2n ${peaks.collect{it.toString()}.sort().join(' ')} \\ | mergeBed -c $mergecols -o $collapsecols > ${prefix}.txt @@ -1262,23 +1262,23 @@ process MERGED_LIB_CONSENSUS { * STEP 6.5: Annotate consensus peaks with HOMER, and add annotation to boolean output file */ process MERGED_LIB_CONSENSUS_ANNOTATE { - label "process_medium" + label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: params.publish_dir_mode when: params.macs_gsize && (replicatesExist || multipleGroups) && !params.skip_consensus_peaks input: - file bed from ch_mlib_macs_consensus_bed - file bool from ch_mlib_macs_consensus_bool - file fasta from ch_fasta - file gtf from ch_gtf + path bed from ch_mlib_macs_consensus_bed + path bool from ch_mlib_macs_consensus_bool + path fasta from ch_fasta + path gtf from ch_gtf output: - file "*.annotatePeaks.txt" + path '*.annotatePeaks.txt' script: - prefix = "consensus_peaks.mLb.clN" + prefix = 'consensus_peaks.mLb.clN' """ annotatePeaks.pl \\ $bed \\ @@ -1304,17 +1304,17 @@ process MERGED_LIB_CONSENSUS_COUNTS { params.macs_gsize && (replicatesExist || multipleGroups) && !params.skip_consensus_peaks input: - file bams from ch_mlib_name_bam_mlib_counts.collect{ it[1] } - file saf from ch_mlib_macs_consensus_saf.collect() + path bams from ch_mlib_name_bam_mlib_counts.collect{ it[1] } + path saf from ch_mlib_macs_consensus_saf.collect() output: - file "*featureCounts.txt" into ch_mlib_macs_consensus_counts - file "*featureCounts.txt.summary" into ch_mlib_macs_consensus_counts_mqc + path '*featureCounts.txt' into ch_mlib_macs_consensus_counts + path '*featureCounts.txt.summary' into ch_mlib_macs_consensus_counts_mqc script: - prefix = "consensus_peaks.mLb.clN" + prefix = 'consensus_peaks.mLb.clN' bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() - pe_params = params.single_end ? '' : "-p --donotsort" + pe_params = params.single_end ? '' : '-p --donotsort' """ featureCounts \\ -F SAF \\ @@ -1335,7 +1335,7 @@ process MERGED_LIB_CONSENSUS_DESEQ2 { label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus/deseq2", mode: params.publish_dir_mode, saveAs: { filename -> - if (filename.endsWith(".igv.txt")) null + if (filename.endsWith('.igv.txt')) null else filename } @@ -1343,22 +1343,22 @@ process MERGED_LIB_CONSENSUS_DESEQ2 { params.macs_gsize && replicatesExist && multipleGroups && !params.skip_consensus_peaks && !params.skip_diff_analysis input: - file counts from ch_mlib_macs_consensus_counts - file mlib_deseq2_pca_header from ch_mlib_deseq2_pca_header - file mlib_deseq2_clustering_header from ch_mlib_deseq2_clustering_header + path counts from ch_mlib_macs_consensus_counts + path mlib_deseq2_pca_header from ch_mlib_deseq2_pca_header + path mlib_deseq2_clustering_header from ch_mlib_deseq2_clustering_header output: - file "*.tsv" into ch_mlib_macs_consensus_deseq_mqc - file "*igv.txt" into ch_mlib_macs_consensus_deseq_comp_igv - file "*.{RData,results.txt,pdf,log}" - file "sizeFactors" - file "*vs*/*.{pdf,txt}" - file "*vs*/*.bed" + path '*.tsv' into ch_mlib_macs_consensus_deseq_mqc + path '*igv.txt' into ch_mlib_macs_consensus_deseq_comp_igv + path '*.{RData,results.txt,pdf,log}' + path 'sizeFactors' + path '*vs*/*.{pdf,txt}' + path '*vs*/*.bed' script: - prefix = "consensus_peaks.mLb.clN" - bam_ext = params.single_end ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" - vst = params.deseq2_vst ? "--vst TRUE" : "" + prefix = 'consensus_peaks.mLb.clN' + bam_ext = params.single_end ? '.mLb.clN.sorted.bam' : '.mLb.clN.bam' + vst = params.deseq2_vst ? '--vst TRUE' : '' """ featurecounts_deseq2.r \\ --featurecount_file $counts \\ @@ -1389,16 +1389,16 @@ process MERGED_LIB_ATAQV { input: tuple val(name), path(bam), path(peak) from ch_mlib_bam_ataqv.join(ch_mlib_macs_ataqv, by: [0]) - file autosomes from ch_genome_autosomes.collect() - file tss_bed from ch_tss_bed + path autosomes from ch_genome_autosomes.collect() + path tss_bed from ch_tss_bed output: - file "*.json" into ch_mlib_ataqv + path '*.json' into ch_mlib_ataqv script: suffix = 'mLb.clN' - peak_param = params.macs_gsize ? "--peak-file ${peak}" : "" - mito_param = params.mito_name ? "--mitochondrial-reference-name ${params.mito_name}" : "" + peak_param = params.macs_gsize ? "--peak-file ${peak}" : '' + mito_param = params.mito_name ? "--mitochondrial-reference-name ${params.mito_name}" : '' """ ataqv \\ --threads $task.cpus \\ @@ -1425,10 +1425,10 @@ process MERGED_LIB_ATAQV_MKARV { !params.skip_ataqv input: - file json from ch_mlib_ataqv.collect() + path json from ch_mlib_ataqv.collect() output: - file "html" + path 'html' script: """ @@ -1462,10 +1462,10 @@ process MERGED_REP_BAM { label 'process_medium' publishDir "${params.outdir}/bwa/mergedReplicate", mode: params.publish_dir_mode, saveAs: { filename -> - if (filename.endsWith(".flagstat")) "samtools_stats/$filename" - else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" - else if (filename.endsWith(".stats")) "samtools_stats/$filename" - else if (filename.endsWith(".metrics.txt")) "picard_metrics/$filename" + if (filename.endsWith('.flagstat')) "samtools_stats/$filename" + else if (filename.endsWith('.idxstats')) "samtools_stats/$filename" + else if (filename.endsWith('.stats')) "samtools_stats/$filename" + else if (filename.endsWith('.metrics.txt')) "picard_metrics/$filename" else filename } @@ -1474,12 +1474,12 @@ process MERGED_REP_BAM { output: tuple val(name), path("*${prefix}.sorted.{bam,bam.bai}") into ch_mrep_bam_bigwig, - ch_mrep_bam_macs - tuple val(name), path("*.flagstat") into ch_mrep_bam_flagstat_bigwig, - ch_mrep_bam_flagstat_macs, - ch_mrep_bam_flagstat_mqc - file "*.{idxstats,stats}" into ch_mrep_bam_stats_mqc - file "*.txt" into ch_mrep_bam_metrics_mqc + ch_mrep_bam_macs + tuple val(name), path('*.flagstat') into ch_mrep_bam_flagstat_bigwig, + ch_mrep_bam_flagstat_macs, + ch_mrep_bam_flagstat_mqc + path '*.{idxstats,stats}' into ch_mrep_bam_stats_mqc + path '*.txt' into ch_mrep_bam_metrics_mqc when: !params.skip_merge_replicates && replicatesExist @@ -1489,7 +1489,7 @@ process MERGED_REP_BAM { bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() def avail_mem = 3 if (!task.memory) { - log.info "[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." + log.info '[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this.' } else { avail_mem = task.memory.toGiga() } @@ -1545,8 +1545,8 @@ process MERGED_REP_BIGWIG { label 'process_medium' publishDir "${params.outdir}/bwa/mergedReplicate/bigwig", mode: params.publish_dir_mode, saveAs: { filename -> - if (filename.endsWith("scale_factor.txt")) "scale/$filename" - else if (filename.endsWith(".bigWig")) "$filename" + if (filename.endsWith('scale_factor.txt')) "scale/$filename" + else if (filename.endsWith('.bigWig')) filename else null } @@ -1555,16 +1555,16 @@ process MERGED_REP_BIGWIG { input: tuple val(name), path(bam), path(flagstat) from ch_mrep_bam_bigwig.join(ch_mrep_bam_flagstat_bigwig, by: [0]) - file sizes from ch_genome_sizes_mrep_bigwig.collect() + path sizes from ch_genome_sizes_mrep_bigwig.collect() output: - tuple val(name), path("*.bigWig") into ch_mrep_bigwig - file "*igv.txt" into ch_mrep_bigwig_igv - file "*scale_factor.txt" + tuple val(name), path('*.bigWig') into ch_mrep_bigwig + path '*igv.txt' into ch_mrep_bigwig_igv + path '*scale_factor.txt' script: prefix = "${name}.mRp.clN" - pe_fragment = params.single_end ? "" : "-pc" + pe_fragment = params.single_end ? '' : '-pc' extend = (params.single_end && params.fragment_size > 0) ? "-fs ${params.fragment_size}" : '' """ SCALE_FACTOR=\$(grep 'mapped (' $flagstat | awk '{print 1000000/\$1}') @@ -1585,8 +1585,8 @@ process MERGED_REP_MACS2 { label 'process_medium' publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}", mode: params.publish_dir_mode, saveAs: { filename -> - if (filename.endsWith(".tsv")) "qc/$filename" - else if (filename.endsWith(".igv.txt")) null + if (filename.endsWith('.tsv')) "qc/$filename" + else if (filename.endsWith('.igv.txt')) null else filename } @@ -1595,22 +1595,22 @@ process MERGED_REP_MACS2 { input: tuple val(name), path(bam), path(flagstat) from ch_mrep_bam_macs.join(ch_mrep_bam_flagstat_macs, by: [0]) - file mrep_peak_count_header from ch_mrep_peak_count_header - file mrep_frip_score_header from ch_mrep_frip_score_header + path mrep_peak_count_header from ch_mrep_peak_count_header + path mrep_frip_score_header from ch_mrep_frip_score_header output: tuple val(name), path("*$PEAK_TYPE") into ch_mrep_macs_homer, - ch_mrep_macs_qc, - ch_mrep_macs_consensus - file "*igv.txt" into ch_mrep_macs_igv - file "*_mqc.tsv" into ch_mrep_macs_mqc - path "*.{bed,xls,gappedPeak,bdg}" + ch_mrep_macs_qc, + ch_mrep_macs_consensus + path '*igv.txt' into ch_mrep_macs_igv + path '*_mqc.tsv' into ch_mrep_macs_mqc + path '*.{bed,xls,gappedPeak,bdg}' script: prefix = "${name}.mRp.clN" broad = params.narrow_peak ? '' : "--broad --broad-cutoff ${params.broad_cutoff}" - format = params.single_end ? "BAM" : "BAMPE" - pileup = params.save_macs_pileup ? "-B --SPMR" : "" + format = params.single_end ? 'BAM' : 'BAMPE' + pileup = params.save_macs_pileup ? '-B --SPMR' : '' """ macs2 callpeak \\ -t ${bam[0]} \\ @@ -1636,7 +1636,7 @@ process MERGED_REP_MACS2 { */ process MERGED_REP_MACS2_ANNOTATE { tag "$name" - label "process_medium" + label 'process_medium' publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}", mode: params.publish_dir_mode when: @@ -1644,11 +1644,11 @@ process MERGED_REP_MACS2_ANNOTATE { input: tuple val(name), path(peak) from ch_mrep_macs_homer - file fasta from ch_fasta - file gtf from ch_gtf + path fasta from ch_fasta + path gtf from ch_gtf output: - file "*.txt" into ch_mrep_macs_annotate + path '*.txt' into ch_mrep_macs_annotate script: prefix = "${name}.mRp.clN" @@ -1667,20 +1667,20 @@ process MERGED_REP_MACS2_ANNOTATE { * STEP 8.4: Aggregated QC plots for peaks, FRiP and peak-to-gene annotation */ process MERGED_REP_MACS2_QC { - label "process_medium" + label 'process_medium' publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/qc", mode: params.publish_dir_mode when: !params.skip_merge_replicates && replicatesExist && params.macs_gsize && !params.skip_peak_qc input: - file peaks from ch_mrep_macs_qc.collect{ it[1] } - file annos from ch_mrep_macs_annotate.collect() - file mrep_peak_annotation_header from ch_mrep_peak_annotation_header + path peaks from ch_mrep_macs_qc.collect{ it[1] } + path annos from ch_mrep_macs_annotate.collect() + path mrep_peak_annotation_header from ch_mrep_peak_annotation_header output: - file "*.tsv" into ch_mrep_peak_qc_mqc - file "*.{txt,pdf}" + path '*.tsv' into ch_mrep_peak_qc_mqc + path '*.{txt,pdf}' script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ suffix = 'mRp.clN' @@ -1708,7 +1708,7 @@ process MERGED_REP_CONSENSUS { label 'process_long' publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: params.publish_dir_mode, saveAs: { filename -> - if (filename.endsWith(".igv.txt")) null + if (filename.endsWith('.igv.txt')) null else filename } @@ -1716,21 +1716,21 @@ process MERGED_REP_CONSENSUS { !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks input: - file peaks from ch_mrep_macs_consensus.collect{ it[1] } + path peaks from ch_mrep_macs_consensus.collect{ it[1] } output: - file "*.bed" into ch_mrep_macs_consensus_bed - file "*.saf" into ch_mrep_macs_consensus_saf - file "*.boolean.txt" into ch_mrep_macs_consensus_bool - file "*igv.txt" into ch_mrep_macs_consensus_igv - file "*.intersect.{txt,plot.pdf}" + path '*.bed' into ch_mrep_macs_consensus_bed + path '*.saf' into ch_mrep_macs_consensus_saf + path '*.boolean.txt' into ch_mrep_macs_consensus_bool + path '*igv.txt' into ch_mrep_macs_consensus_igv + path '*.intersect.{txt,plot.pdf}' script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ suffix = 'mRp.clN' prefix = "consensus_peaks.${suffix}" mergecols = params.narrow_peak ? (2..10).join(',') : (2..9).join(',') - collapsecols = params.narrow_peak ? (["collapse"]*9).join(',') : (["collapse"]*8).join(',') - expandparam = params.narrow_peak ? "--is_narrow_peak" : "" + collapsecols = params.narrow_peak ? (['collapse']*9).join(',') : (['collapse']*8).join(',') + expandparam = params.narrow_peak ? '--is_narrow_peak' : '' """ sort -T '.' -k1,1 -k2,2n ${peaks.collect{it.toString()}.sort().join(' ')} \\ | mergeBed -c $mergecols -o $collapsecols > ${prefix}.txt @@ -1757,23 +1757,23 @@ process MERGED_REP_CONSENSUS { * STEP 8.6: Annotate consensus peaks with HOMER, and add annotation to boolean output file */ process MERGED_REP_CONSENSUS_ANNOTATE { - label "process_medium" + label 'process_medium' publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: params.publish_dir_mode when: !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks input: - file bed from ch_mrep_macs_consensus_bed - file bool from ch_mrep_macs_consensus_bool - file fasta from ch_fasta - file gtf from ch_gtf + path bed from ch_mrep_macs_consensus_bed + path bool from ch_mrep_macs_consensus_bool + path fasta from ch_fasta + path gtf from ch_gtf output: - file "*.annotatePeaks.txt" + path '*.annotatePeaks.txt' script: - prefix = "consensus_peaks.mRp.clN" + prefix = 'consensus_peaks.mRp.clN' """ annotatePeaks.pl \\ $bed \\ @@ -1799,17 +1799,17 @@ process MERGED_REP_CONSENSUS_COUNTS { !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks input: - file bams from ch_mlib_name_bam_mrep_counts.collect{ it[1] } - file saf from ch_mrep_macs_consensus_saf.collect() + path bams from ch_mlib_name_bam_mrep_counts.collect{ it[1] } + path saf from ch_mrep_macs_consensus_saf.collect() output: - file "*featureCounts.txt" into ch_mrep_macs_consensus_counts - file "*featureCounts.txt.summary" into ch_mrep_macs_consensus_counts_mqc + path '*featureCounts.txt' into ch_mrep_macs_consensus_counts + path '*featureCounts.txt.summary' into ch_mrep_macs_consensus_counts_mqc script: - prefix = "consensus_peaks.mRp.clN" + prefix = 'consensus_peaks.mRp.clN' bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() - pe_params = params.single_end ? '' : "-p --donotsort" + pe_params = params.single_end ? '' : '-p --donotsort' """ featureCounts \\ -F SAF \\ @@ -1830,7 +1830,7 @@ process MERGED_REP_CONSENSUS_DESEQ2 { label 'process_medium' publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus/deseq2", mode: params.publish_dir_mode, saveAs: { filename -> - if (filename.endsWith(".igv.txt")) null + if (filename.endsWith('.igv.txt')) null else filename } @@ -1838,22 +1838,22 @@ process MERGED_REP_CONSENSUS_DESEQ2 { !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks && !params.skip_diff_analysis input: - file counts from ch_mrep_macs_consensus_counts - file mrep_deseq2_pca_header from ch_mrep_deseq2_pca_header - file mrep_deseq2_clustering_header from ch_mrep_deseq2_clustering_header + path counts from ch_mrep_macs_consensus_counts + path mrep_deseq2_pca_header from ch_mrep_deseq2_pca_header + path mrep_deseq2_clustering_header from ch_mrep_deseq2_clustering_header output: - file "*.tsv" into ch_mrep_macs_consensus_deseq_mqc - file "*igv.txt" into ch_mrep_macs_consensus_deseq_comp_igv - file "*.{RData,results.txt,pdf,log}" - file "sizeFactors" - file "*vs*/*.{pdf,txt}" - file "*vs*/*.bed" + path '*.tsv' into ch_mrep_macs_consensus_deseq_mqc + path '*igv.txt' into ch_mrep_macs_consensus_deseq_comp_igv + path '*.{RData,results.txt,pdf,log}' + path 'sizeFactors' + path '*vs*/*.{pdf,txt}' + path '*vs*/*.bed' script: - prefix = "consensus_peaks.mRp.clN" - bam_ext = params.single_end ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" - vst = params.deseq2_vst ? "--vst TRUE" : "" + prefix = 'consensus_peaks.mRp.clN' + bam_ext = params.single_end ? '.mLb.clN.sorted.bam' : '.mLb.clN.bam' + vst = params.deseq2_vst ? '--vst TRUE' : '' """ featurecounts_deseq2.r \\ --featurecount_file $counts \\ @@ -1889,20 +1889,20 @@ process IGV { !params.skip_igv input: - file fasta from ch_fasta + path fasta from ch_fasta - file bigwigs from ch_mlib_bigwig_igv.collect().ifEmpty([]) - file peaks from ch_mlib_macs_igv.collect().ifEmpty([]) - file consensus_peaks from ch_mlib_macs_consensus_igv.collect().ifEmpty([]) - file differential_peaks from ch_mlib_macs_consensus_deseq_comp_igv.collect().ifEmpty([]) + path bigwigs from ch_mlib_bigwig_igv.collect().ifEmpty([]) + path peaks from ch_mlib_macs_igv.collect().ifEmpty([]) + path consensus_peaks from ch_mlib_macs_consensus_igv.collect().ifEmpty([]) + path differential_peaks from ch_mlib_macs_consensus_deseq_comp_igv.collect().ifEmpty([]) - file rbigwigs from ch_mrep_bigwig_igv.collect().ifEmpty([]) - file rpeaks from ch_mrep_macs_igv.collect().ifEmpty([]) - file rconsensus_peaks from ch_mrep_macs_consensus_igv.collect().ifEmpty([]) - file rdifferential_peaks from ch_mrep_macs_consensus_deseq_comp_igv.collect().ifEmpty([]) + path rbigwigs from ch_mrep_bigwig_igv.collect().ifEmpty([]) + path rpeaks from ch_mrep_macs_igv.collect().ifEmpty([]) + path rconsensus_peaks from ch_mrep_macs_consensus_igv.collect().ifEmpty([]) + path rdifferential_peaks from ch_mrep_macs_consensus_deseq_comp_igv.collect().ifEmpty([]) output: - file "*.{txt,xml}" + path '*.{txt,xml}' script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ """ @@ -1925,13 +1925,13 @@ process IGV { process get_software_versions { publishDir "${params.outdir}/pipeline_info", mode: params.publish_dir_mode, saveAs: { filename -> - if (filename.indexOf(".csv") > 0) filename + if (filename.indexOf('.csv') > 0) filename else null } output: - file 'software_versions_mqc.yaml' into ch_software_versions_mqc - file "software_versions.csv" + path 'software_versions_mqc.yaml' into ch_software_versions_mqc + path 'software_versions.csv' script: """ @@ -1983,42 +1983,42 @@ process MULTIQC { !params.skip_multiqc input: - file (multiqc_config) from ch_multiqc_config - file (mqc_custom_config) from ch_multiqc_custom_config.collect().ifEmpty([]) - - file ('software_versions/*') from ch_software_versions_mqc.collect() - file workflow_summary from ch_workflow_summary.collectFile(name: "workflow_summary_mqc.yaml") - - file ('fastqc/*') from ch_fastqc_reports_mqc.collect().ifEmpty([]) - file ('trimgalore/*') from ch_trimgalore_results_mqc.collect().ifEmpty([]) - file ('trimgalore/fastqc/*') from ch_trimgalore_fastqc_reports_mqc.collect().ifEmpty([]) - - file ('alignment/library/*') from ch_sort_bam_flagstat_mqc.collect() - - file ('alignment/mergedLibrary/*') from ch_mlib_bam_stats_mqc.collect() - file ('alignment/mergedLibrary/*') from ch_mlib_rm_orphan_flagstat_mqc.collect{it[1]} - file ('alignment/mergedLibrary/*') from ch_mlib_rm_orphan_stats_mqc.collect() - file ('alignment/mergedLibrary/picard_metrics/*') from ch_mlib_bam_metrics_mqc.collect() - file ('alignment/mergedLibrary/picard_metrics/*') from ch_mlib_collectmetrics_mqc.collect() - file ('macs/mergedLibrary/*') from ch_mlib_macs_mqc.collect().ifEmpty([]) - file ('macs/mergedLibrary/*') from ch_mlib_peak_qc_mqc.collect().ifEmpty([]) - file ('macs/mergedLibrary/consensus/*') from ch_mlib_macs_consensus_counts_mqc.collect().ifEmpty([]) - file ('macs/mergedLibrary/consensus/*') from ch_mlib_macs_consensus_deseq_mqc.collect().ifEmpty([]) - file ('preseq/*') from ch_mlib_preseq_mqc.collect().ifEmpty([]) - file ('deeptools/*') from ch_mlib_plotprofile_mqc.collect().ifEmpty([]) - file ('deeptools/*') from ch_mlib_plotfingerprint_mqc.collect().ifEmpty([]) - - file ('alignment/mergedReplicate/*') from ch_mrep_bam_flagstat_mqc.collect{it[1]}.ifEmpty([]) - file ('alignment/mergedReplicate/*') from ch_mrep_bam_stats_mqc.collect().ifEmpty([]) - file ('alignment/mergedReplicate/*') from ch_mrep_bam_metrics_mqc.collect().ifEmpty([]) - file ('macs/mergedReplicate/*') from ch_mrep_macs_mqc.collect().ifEmpty([]) - file ('macs/mergedReplicate/*') from ch_mrep_peak_qc_mqc.collect().ifEmpty([]) - file ('macs/mergedReplicate/consensus/*') from ch_mrep_macs_consensus_counts_mqc.collect().ifEmpty([]) - file ('macs/mergedReplicate/consensus/*') from ch_mrep_macs_consensus_deseq_mqc.collect().ifEmpty([]) + path (multiqc_config) from ch_multiqc_config + path (mqc_custom_config) from ch_multiqc_custom_config.collect().ifEmpty([]) + + path ('software_versions/*') from ch_software_versions_mqc.collect() + path workflow_summary from ch_workflow_summary.collectFile(name: 'workflow_summary_mqc.yaml') + + path ('fastqc/*') from ch_fastqc_reports_mqc.collect().ifEmpty([]) + path ('trimgalore/*') from ch_trimgalore_results_mqc.collect().ifEmpty([]) + path ('trimgalore/fastqc/*') from ch_trimgalore_fastqc_reports_mqc.collect().ifEmpty([]) + + path ('alignment/library/*') from ch_sort_bam_flagstat_mqc.collect() + + path ('alignment/mergedLibrary/*') from ch_mlib_bam_stats_mqc.collect() + path ('alignment/mergedLibrary/*') from ch_mlib_rm_orphan_flagstat_mqc.collect{it[1]} + path ('alignment/mergedLibrary/*') from ch_mlib_rm_orphan_stats_mqc.collect() + path ('alignment/mergedLibrary/picard_metrics/*') from ch_mlib_bam_metrics_mqc.collect() + path ('alignment/mergedLibrary/picard_metrics/*') from ch_mlib_collectmetrics_mqc.collect() + path ('macs/mergedLibrary/*') from ch_mlib_macs_mqc.collect().ifEmpty([]) + path ('macs/mergedLibrary/*') from ch_mlib_peak_qc_mqc.collect().ifEmpty([]) + path ('macs/mergedLibrary/consensus/*') from ch_mlib_macs_consensus_counts_mqc.collect().ifEmpty([]) + path ('macs/mergedLibrary/consensus/*') from ch_mlib_macs_consensus_deseq_mqc.collect().ifEmpty([]) + path ('preseq/*') from ch_mlib_preseq_mqc.collect().ifEmpty([]) + path ('deeptools/*') from ch_mlib_plotprofile_mqc.collect().ifEmpty([]) + path ('deeptools/*') from ch_mlib_plotfingerprint_mqc.collect().ifEmpty([]) + + path ('alignment/mergedReplicate/*') from ch_mrep_bam_flagstat_mqc.collect{it[1]}.ifEmpty([]) + path ('alignment/mergedReplicate/*') from ch_mrep_bam_stats_mqc.collect().ifEmpty([]) + path ('alignment/mergedReplicate/*') from ch_mrep_bam_metrics_mqc.collect().ifEmpty([]) + path ('macs/mergedReplicate/*') from ch_mrep_macs_mqc.collect().ifEmpty([]) + path ('macs/mergedReplicate/*') from ch_mrep_peak_qc_mqc.collect().ifEmpty([]) + path ('macs/mergedReplicate/consensus/*') from ch_mrep_macs_consensus_counts_mqc.collect().ifEmpty([]) + path ('macs/mergedReplicate/consensus/*') from ch_mrep_macs_consensus_deseq_mqc.collect().ifEmpty([]) output: - file "*multiqc_report.html" into ch_multiqc_report - file "*_data" + path '*multiqc_report.html' into ch_multiqc_report + path '*_data' script: rtitle = custom_runName ? "--title \"$custom_runName\"" : '' @@ -2044,11 +2044,11 @@ process output_documentation { publishDir "${params.outdir}/Documentation", mode: params.publish_dir_mode input: - file output_docs from ch_output_docs - file images from ch_output_docs_images + path output_docs from ch_output_docs + path images from ch_output_docs_images output: - file "results_description.html" + path 'results_description.html' script: """ From 3a73c81f9c040fb44a840ab67069792536efee0c Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 2 Jun 2020 11:23:34 +0100 Subject: [PATCH 182/226] Update AWS tests --- .github/workflows/awstest.yml | 10 ++++++++-- 1 file changed, 8 insertions(+), 2 deletions(-) diff --git a/.github/workflows/awstest.yml b/.github/workflows/awstest.yml index 8ccdb4cf..af704a87 100644 --- a/.github/workflows/awstest.yml +++ b/.github/workflows/awstest.yml @@ -11,6 +11,7 @@ on: jobs: run-awstest: + if: github.repository == 'nf-core/atacseq' name: Run AWS test runs-on: ubuntu-latest steps: @@ -26,5 +27,10 @@ jobs: AWS_ACCESS_KEY_ID: ${{secrets.AWS_KEY_ID}} AWS_SECRET_ACCESS_KEY: ${{secrets.AWS_KEY_SECRET}} TOWER_ACCESS_TOKEN: ${{secrets.TOWER_ACCESS_TOKEN}} - run: | - aws batch submit-job --region eu-west-1 --job-name nf-core-atacseq --job-queue 'default-8b3836e0-5eda-11ea-96e5-0a2c3f6a2a32' --job-definition nextflow --container-overrides '{"command": ["nf-core/atacseq", "-r '"${GITHUB_SHA}"' -profile test_full --outdir s3://nf-core-awsmegatests/atacseq/results-'"${GITHUB_SHA}"' -w s3://nf-core-awsmegatests/atacseq/work-'"${GITHUB_SHA}"' -with-tower"], "environment": [{"name": "TOWER_ACCESS_TOKEN", "value": "'"$TOWER_ACCESS_TOKEN"'"}]}' + run: | # Submits job to AWS batch using a 'nextflow-4GiB' instance. Setting JVM options to "-XX:+UseG1GC" for more efficient garbage collection when staging remote files. + aws batch submit-job \ + --region eu-west-1 \ + --job-name nf-core-atacseq \ + --job-queue 'default-8b3836e0-5eda-11ea-96e5-0a2c3f6a2a32' \ + --job-definition nextflow-4GiB \ + --container-overrides '{"command": ["nf-core/atacseq", "-r '"${GITHUB_SHA}"' -profile test_full --outdir s3://nf-core-awsmegatests/atacseq/results-'"${GITHUB_SHA}"' -w s3://nf-core-awsmegatests/atacseq/work-'"${GITHUB_SHA}"' -with-tower"], "environment": [{"name": "TOWER_ACCESS_TOKEN", "value": "'"$TOWER_ACCESS_TOKEN"'"}, {"name": "NXF_OPTS", "value": "-XX:+UseG1GC"}]}' From b4600599f9ec3911becf01f5a2d38138449ea0fe Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 2 Jun 2020 13:48:27 +0100 Subject: [PATCH 183/226] Fix chipseq #154 --- CHANGELOG.md | 1 + docs/output.md | 2 +- main.nf | 4 ++-- 3 files changed, 4 insertions(+), 3 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index c147c815..b905a7ce 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -29,6 +29,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * [#102](https://github.com/nf-core/atacseq/issues/102) - Incorrect Group ID assigned by featurecounts_deseq2.r * [nf-core/chipseq#118](https://github.com/nf-core/chipseq/issues/118) - Running on with SGE * [nf-core/chipseq#132](https://github.com/nf-core/chipseq/issues/132) - BigWig Error: sort: cannot create temporary file in '': Read-only file system +* [nf-core/chipseq#154](https://github.com/nf-core/chipseq/issues/154) - computeMatrix.val.mat.gz files not zipped * Make executables in `bin/` compatible with Python 3 ### `Dependencies` diff --git a/docs/output.md b/docs/output.md index b1a4a71c..a2f40add 100755 --- a/docs/output.md +++ b/docs/output.md @@ -140,7 +140,7 @@ The library-level alignments associated with the same sample are merged and subs * `bwa/mergedLibrary/deepTools/plotFingerprint/` * Output files: `*.plotFingerprint.pdf`, `*.plotFingerprint.qcmetrics.txt`, `*.plotFingerprint.raw.txt` * `bwa/mergedLibrary/deepTools/plotProfile/` - * Output files: `*.computeMatrix.mat.gz`, `*.computeMatrix.vals.mat.gz`, `*.plotProfile.pdf`, `*.plotProfile.tab`. + * Output files: `*.computeMatrix.mat.gz`, `*.computeMatrix.vals.mat.tab`, `*.plotProfile.pdf`, `*.plotProfile.tab`. 4. **Call peaks** diff --git a/main.nf b/main.nf index 82fb4ee4..54f006cc 100644 --- a/main.nf +++ b/main.nf @@ -1014,7 +1014,7 @@ process MERGED_LIB_PLOTPROFILE { output: path '*.plotProfile.tab' into ch_mlib_plotprofile_mqc - path '*.{gz,pdf}' + path '*.{gz,pdf,mat.tab}' script: prefix = "${name}.mLb.clN" @@ -1023,7 +1023,7 @@ process MERGED_LIB_PLOTPROFILE { --regionsFileName $bed \\ --scoreFileName $bigwig \\ --outFileName ${prefix}.computeMatrix.mat.gz \\ - --outFileNameMatrix ${prefix}.computeMatrix.vals.mat.gz \\ + --outFileNameMatrix ${prefix}.computeMatrix.vals.mat.tab \\ --regionBodyLength 1000 \\ --beforeRegionStartLength 3000 \\ --afterRegionStartLength 3000 \\ From b12507c242a428687ee7ab755ebabac51b5929bb Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 2 Jun 2020 16:14:43 +0100 Subject: [PATCH 184/226] Add --skip_peak_annotation parameter --- CHANGELOG.md | 1 + docs/usage.md | 5 +++++ main.nf | 14 ++++++++------ nextflow.config | 1 + 4 files changed, 15 insertions(+), 6 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index b905a7ce..d563c140 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -18,6 +18,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * Capitalise process names * Parameters: * `--skip_peak_qc` to skip MACS2 peak QC plot generation + * `--skip_peak_annotation` to skip annotation of MACS2 and consensus peaks with HOMER * `--skip_consensus_peaks` to skip consensus peak generation * `--deseq2_vst` to use variance stabilizing transformation (VST) instead of regularized log transformation (rlog) with DESeq2 * `--publish_dir_mode` to customise method of publishing results to output directory [nf-core/tools#585](https://github.com/nf-core/tools/issues/585) diff --git a/docs/usage.md b/docs/usage.md index ec64225c..b8252113 100755 --- a/docs/usage.md +++ b/docs/usage.md @@ -42,6 +42,7 @@ * [`--min_reps_consensus`](#--min_reps_consensus) * [`--save_macs_pileup`](#--save_macs_pileup) * [`--skip_peak_qc`](#--skip_peak_qc) + * [`--skip_peak_annotation`](#--skip_peak_annotation) * [`--skip_consensus_peaks`](#--skip_consensus_peaks) * [Differential analysis](#differential_analysis) * [`--deseq2_vst`](#--deseq2_vst) @@ -403,6 +404,10 @@ Instruct MACS2 to create bedGraph files using the `-B --SPMR` parameters. Skip MACS2 peak QC plot generation. +### `--skip_peak_annotation` + +Skip annotation of MACS2 and consensus peaks with HOMER. + ### `--skip_consensus_peaks` Skip consensus peak generation, annotation and counting. diff --git a/main.nf b/main.nf index 54f006cc..5f57775c 100644 --- a/main.nf +++ b/main.nf @@ -63,6 +63,7 @@ def helpMessage() { --min_reps_consensus [int] Number of biological replicates required from a given condition for a peak to contribute to a consensus peak (Default: 1) --save_macs_pileup [bool] Instruct MACS2 to create bedGraph files normalised to signal per million reads --skip_peak_qc [bool] Skip MACS2 peak QC plot generation + --skip_peak_annotation [bool] Skip annotation of MACS2 and consensus peaks with HOMER --skip_consensus_peaks [bool] Skip consensus peak generation Differential analysis @@ -261,6 +262,7 @@ if (params.save_align_intermeds) summary['Save Intermeds'] = 'Yes' if (params.save_macs_pileup) summary['Save MACS2 Pileup'] = 'Yes' if (params.skip_merge_replicates) summary['Skip Merge Replicates'] = 'Yes' if (params.skip_peak_qc) summary['Skip MACS2 Peak QC'] = 'Yes' +if (params.skip_peak_annotation) summary['Skip Peak Annotation'] = 'Yes' if (params.skip_consensus_peaks) summary['Skip Consensus Peaks'] = 'Yes' if (params.deseq2_vst) summary['Use DESeq2 vst Transform'] = 'Yes' if (params.skip_diff_analysis) summary['Skip Differential Analysis'] = 'Yes' @@ -1144,7 +1146,7 @@ process MERGED_LIB_MACS2_ANNOTATE { publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}", mode: params.publish_dir_mode when: - params.macs_gsize + params.macs_gsize && !params.skip_peak_annotation input: tuple val(name), path(peak) from ch_mlib_macs_homer @@ -1175,7 +1177,7 @@ process MERGED_LIB_MACS2_QC { publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/qc", mode: params.publish_dir_mode when: - params.macs_gsize && !params.skip_peak_qc + params.macs_gsize && !params.skip_peak_annotation && !params.skip_peak_qc input: path peaks from ch_mlib_macs_qc.collect{ it[1] } @@ -1266,7 +1268,7 @@ process MERGED_LIB_CONSENSUS_ANNOTATE { publishDir "${params.outdir}/bwa/mergedLibrary/macs/${PEAK_TYPE}/consensus", mode: params.publish_dir_mode when: - params.macs_gsize && (replicatesExist || multipleGroups) && !params.skip_consensus_peaks + params.macs_gsize && (replicatesExist || multipleGroups) && !params.skip_consensus_peaks && !params.skip_peak_annotation input: path bed from ch_mlib_macs_consensus_bed @@ -1640,7 +1642,7 @@ process MERGED_REP_MACS2_ANNOTATE { publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}", mode: params.publish_dir_mode when: - !params.skip_merge_replicates && replicatesExist && params.macs_gsize + !params.skip_merge_replicates && replicatesExist && params.macs_gsize && !params.skip_peak_annotation input: tuple val(name), path(peak) from ch_mrep_macs_homer @@ -1671,7 +1673,7 @@ process MERGED_REP_MACS2_QC { publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/qc", mode: params.publish_dir_mode when: - !params.skip_merge_replicates && replicatesExist && params.macs_gsize && !params.skip_peak_qc + !params.skip_merge_replicates && replicatesExist && params.macs_gsize && !params.skip_peak_qc && !params.skip_peak_annotation input: path peaks from ch_mrep_macs_qc.collect{ it[1] } @@ -1761,7 +1763,7 @@ process MERGED_REP_CONSENSUS_ANNOTATE { publishDir "${params.outdir}/bwa/mergedReplicate/macs/${PEAK_TYPE}/consensus", mode: params.publish_dir_mode when: - !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks + !params.skip_merge_replicates && replicatesExist && params.macs_gsize && multipleGroups && !params.skip_consensus_peaks && !params.skip_peak_annotation input: path bed from ch_mrep_macs_consensus_bed diff --git a/nextflow.config b/nextflow.config index e0adef1b..aafd8841 100644 --- a/nextflow.config +++ b/nextflow.config @@ -42,6 +42,7 @@ params { min_reps_consensus = 1 save_macs_pileup = false skip_peak_qc = false + skip_peak_annotation = false skip_consensus_peaks = false // Options: Differential analysis From 4af0fbb5a1b1fcb4d35093e9615c97cc70d2424d Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 2 Jun 2020 16:25:57 +0100 Subject: [PATCH 185/226] Add default values --- main.nf | 80 ++++++++++++++++++++++++++++----------------------------- 1 file changed, 40 insertions(+), 40 deletions(-) diff --git a/main.nf b/main.nf index 5f57775c..624921a8 100644 --- a/main.nf +++ b/main.nf @@ -19,27 +19,27 @@ def helpMessage() { nextflow run nf-core/atacseq --input design.csv --genome GRCh37 -profile docker Mandatory arguments: - --input [file] Comma-separated file containing information about the samples in the experiment (see docs/usage.md) - --fasta [file] Path to Fasta reference. Not mandatory when using reference in iGenomes config via --genome - --gtf [file] Path to GTF file. Not mandatory when using reference in iGenomes config via --genome + --input [file] Comma-separated file containing information about the samples in the experiment (see docs/usage.md) (Default: './design.csv') + --fasta [file] Path to Fasta reference. Not mandatory when using reference in iGenomes config via --genome (Default: false) + --gtf [file] Path to GTF file. Not mandatory when using reference in iGenomes config via --genome (Default: false) -profile [str] Configuration profile to use. Can use multiple (comma separated) Available: conda, docker, singularity, awsbatch, test Generic - --single_end [bool] Specifies that the input is single-end reads - --seq_center [str] Sequencing center information to be added to read group of BAM files + --single_end [bool] Specifies that the input is single-end reads (Default: false) + --seq_center [str] Sequencing center information to be added to read group of BAM files (Default: false) --fragment_size [int] Estimated fragment size used to extend single-end reads (Default: 0) --fingerprint_bins [int] Number of genomic bins to use when calculating fingerprint plot (Default: 500000) References If not specified in the configuration file or you wish to overwrite any of the references - --genome [str] Name of iGenomes reference - --bwa_index [file] Full path to directory containing BWA index including base name i.e. /path/to/index/genome.fa - --gene_bed [file] Path to BED file containing gene intervals - --tss_bed [file] Path to BED file containing transcription start sites - --macs_gsize [str] Effective genome size parameter required by MACS2. If using iGenomes config, values have only been provided when --genome is set as GRCh37, GRCm38, hg19, mm10, BDGP6 and WBcel235 - --blacklist [file] Path to blacklist regions (.BED format), used for filtering alignments - --mito_name [str] Name of Mitochondrial chomosome in genome fasta (e.g. chrM). Reads aligning to this contig are filtered out - --save_reference [bool] If generated by the pipeline save the BWA index in the results directory + --genome [str] Name of iGenomes reference (Default: false) + --bwa_index [file] Full path to directory containing BWA index including base name i.e. /path/to/index/genome.fa (Default: false) + --gene_bed [file] Path to BED file containing gene intervals (Default: false) + --tss_bed [file] Path to BED file containing transcription start sites (Default: false) + --macs_gsize [str] Effective genome size parameter required by MACS2. If using iGenomes config, values have only been provided when --genome is set as GRCh37, GRCm38, hg19, mm10, BDGP6 and WBcel235 (Default: false) + --blacklist [file] Path to blacklist regions (.BED format), used for filtering alignments (Default: false) + --mito_name [str] Name of Mitochondrial chomosome in genome fasta (e.g. chrM). Reads aligning to this contig are filtered out (Default: false) + --save_reference [bool] If generated by the pipeline save the BWA index in the results directory (Default: false) Trimming --clip_r1 [int] Instructs Trim Galore to remove bp from the 5' end of read 1 (or single-end reads) (Default: 0) @@ -47,46 +47,46 @@ def helpMessage() { --three_prime_clip_r1 [int] Instructs Trim Galore to remove bp from the 3' end of read 1 AFTER adapter/quality trimming has been performed (Default: 0) --three_prime_clip_r2 [int] Instructs Trim Galore to remove bp from the 3' end of read 2 AFTER adapter/quality trimming has been performed (Default: 0) --trim_nextseq [int] Instructs Trim Galore to apply the --nextseq=X option, to trim based on quality after removing poly-G tails (Default: 0) - --skip_trimming [bool] Skip the adapter trimming step - --save_trimmed [bool] Save the trimmed FastQ files in the results directory + --skip_trimming [bool] Skip the adapter trimming step (Default: false) + --save_trimmed [bool] Save the trimmed FastQ files in the results directory (Default: false) Alignments - --keep_mito [bool] Reads mapping to mitochondrial contig are not filtered from alignments - --keep_dups [bool] Duplicate reads are not filtered from alignments - --keep_multi_map [bool] Reads mapping to multiple locations are not filtered from alignments - --skip_merge_replicates [bool] Do not perform alignment merging and downstream analysis by merging replicates i.e. only do this by merging resequenced libraries - --save_align_intermeds [bool] Save the intermediate BAM files from the alignment step - not done by default + --keep_mito [bool] Reads mapping to mitochondrial contig are not filtered from alignments (Default: false) + --keep_dups [bool] Duplicate reads are not filtered from alignments (Default: false) + --keep_multi_map [bool] Reads mapping to multiple locations are not filtered from alignments (Default: false) + --skip_merge_replicates [bool] Do not perform alignment merging and downstream analysis by merging replicates i.e. only do this by merging resequenced libraries (Default: false) + --save_align_intermeds [bool] Save the intermediate BAM files from the alignment step - not done by default (Default: false) Peaks - --narrow_peak [bool] Run MACS2 in narrowPeak mode + --narrow_peak [bool] Run MACS2 in narrowPeak mode (Default: false) --broad_cutoff [float] Specifies broad cutoff value for MACS2. Only used when --narrow_peak isnt specified (Default: 0.1) --min_reps_consensus [int] Number of biological replicates required from a given condition for a peak to contribute to a consensus peak (Default: 1) - --save_macs_pileup [bool] Instruct MACS2 to create bedGraph files normalised to signal per million reads - --skip_peak_qc [bool] Skip MACS2 peak QC plot generation - --skip_peak_annotation [bool] Skip annotation of MACS2 and consensus peaks with HOMER - --skip_consensus_peaks [bool] Skip consensus peak generation + --save_macs_pileup [bool] Instruct MACS2 to create bedGraph files normalised to signal per million reads (Default: false) + --skip_peak_qc [bool] Skip MACS2 peak QC plot generation (Default: false) + --skip_peak_annotation [bool] Skip annotation of MACS2 and consensus peaks with HOMER (Default: false) + --skip_consensus_peaks [bool] Skip consensus peak generation (Default: false) Differential analysis - --deseq2_vst [bool] Use vst transformation instead of rlog with DESeq2 - --skip_diff_analysis [bool] Skip differential accessibility analysis + --deseq2_vst [bool] Use vst transformation instead of rlog with DESeq2 (Default: false) + --skip_diff_analysis [bool] Skip differential accessibility analysis (Default: false) QC - --skip_fastqc [bool] Skip FastQC - --skip_picard_metrics [bool] Skip Picard CollectMultipleMetrics - --skip_preseq [bool] Skip Preseq - --skip_plot_profile [bool] Skip deepTools plotProfile - --skip_plot_fingerprint [bool] Skip deepTools plotFingerprint - --skip_ataqv [bool] Skip Ataqv - --skip_igv [bool] Skip IGV - --skip_multiqc [bool] Skip MultiQC + --skip_fastqc [bool] Skip FastQC (Default: false) + --skip_picard_metrics [bool] Skip Picard CollectMultipleMetrics (Default: false) + --skip_preseq [bool] Skip Preseq (Default: false) + --skip_plot_profile [bool] Skip deepTools plotProfile (Default: false) + --skip_plot_fingerprint [bool] Skip deepTools plotFingerprint (Default: false) + --skip_ataqv [bool] Skip Ataqv (Default: false) + --skip_igv [bool] Skip IGV (Default: false) + --skip_multiqc [bool] Skip MultiQC (Default: false) Other - --outdir [file] The output directory where the results will be saved + --outdir [file] The output directory where the results will be saved (Default: './results') --publish_dir_mode [str] Mode for publishing results in the output directory. Available: symlink, rellink, link, copy, copyNoFollow, move (Default: copy) - --email [email] Set this parameter to your e-mail address to get a summary e-mail with details of the run sent to you when the workflow exits - --email_on_fail [email] Same as --email, except only send mail if the workflow is not successful - --max_multiqc_email_size [str] Theshold size for MultiQC report to be attached in notification email. If file generated by pipeline exceeds the threshold, it will not be attached (Default: 25MB) - -name [str] Name for the pipeline run. If not specified, Nextflow will automatically generate a random mnemonic + --email [email] Set this parameter to your e-mail address to get a summary e-mail with details of the run sent to you when the workflow exits (Default: false) + --email_on_fail [email] Same as --email, except only send mail if the workflow is not successful (Default: false) + --max_multiqc_email_size [str] Threshold size for MultiQC report to be attached in notification email. If file generated by pipeline exceeds the threshold, it will not be attached (Default: 25MB) + -name [str] Name for the pipeline run. If not specified, Nextflow will automatically generate a random mnemonic (Default: false) AWSBatch --awsqueue [str] The AWSBatch JobQueue that needs to be set when running on AWSBatch From 892994c669c7a2bd8e6208f0bcf976118a426926 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 2 Jun 2020 17:01:57 +0100 Subject: [PATCH 186/226] Add plotHeatmap --- docs/output.md | 2 +- main.nf | 6 +++++- 2 files changed, 6 insertions(+), 2 deletions(-) diff --git a/docs/output.md b/docs/output.md index a2f40add..417be661 100755 --- a/docs/output.md +++ b/docs/output.md @@ -140,7 +140,7 @@ The library-level alignments associated with the same sample are merged and subs * `bwa/mergedLibrary/deepTools/plotFingerprint/` * Output files: `*.plotFingerprint.pdf`, `*.plotFingerprint.qcmetrics.txt`, `*.plotFingerprint.raw.txt` * `bwa/mergedLibrary/deepTools/plotProfile/` - * Output files: `*.computeMatrix.mat.gz`, `*.computeMatrix.vals.mat.tab`, `*.plotProfile.pdf`, `*.plotProfile.tab`. + * Output files: `*.computeMatrix.mat.gz`, `*.computeMatrix.vals.mat.tab`, `*.plotProfile.pdf`, `*.plotProfile.tab`, `*.plotHeatmap.pdf`, `*.plotHeatmap.mat.tab`. 4. **Call peaks** diff --git a/main.nf b/main.nf index 624921a8..1f040e2a 100644 --- a/main.nf +++ b/main.nf @@ -1000,7 +1000,7 @@ process MERGED_LIB_BIGWIG { } /* - * STEP 5.4: Generate gene body coverage plot with deepTools plotProfile + * STEP 5.4: Generate gene body coverage plot with deepTools plotProfile and plotHeatmap */ process MERGED_LIB_PLOTPROFILE { tag "$name" @@ -1036,6 +1036,10 @@ process MERGED_LIB_PLOTPROFILE { plotProfile --matrixFile ${prefix}.computeMatrix.mat.gz \\ --outFileName ${prefix}.plotProfile.pdf \\ --outFileNameData ${prefix}.plotProfile.tab + + plotHeatmap --matrixFile ${prefix}.computeMatrix.mat.gz \\ + --outFileName ${prefix}.plotHeatmap.pdf \\ + --outFileNameMatrix ${prefix}.plotHeatmap.mat.tab """ } From 77444f5aaa1ab6bc36ff713d94dacdb0f27395cc Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 2 Jun 2020 17:09:31 +0100 Subject: [PATCH 187/226] Update CHANGELOG --- CHANGELOG.md | 2 ++ 1 file changed, 2 insertions(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index d563c140..7d4aba66 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -14,7 +14,9 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * [#79](https://github.com/nf-core/atacseq/issues/79) - Parallelize DESeq2 * [#80](https://github.com/nf-core/atacseq/pull/80) - Added social preview image * [#97](https://github.com/nf-core/atacseq/issues/97) - PBC1, PBC2 from pipeline? +* [nf-core/chipseq#153](https://github.com/nf-core/chipseq/issues/153) - Add plotHeatmap * Update template to tools `1.9` +* Replace `set` with `tuple` and `file()` with `path()` in all processes * Capitalise process names * Parameters: * `--skip_peak_qc` to skip MACS2 peak QC plot generation From bab37700defd89c4897f0f982f03250b887642f2 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 3 Jun 2020 09:46:27 +0100 Subject: [PATCH 188/226] Update AWS tests --- .github/workflows/awstest.yml | 1 + 1 file changed, 1 insertion(+) diff --git a/.github/workflows/awstest.yml b/.github/workflows/awstest.yml index af704a87..c315d213 100644 --- a/.github/workflows/awstest.yml +++ b/.github/workflows/awstest.yml @@ -6,6 +6,7 @@ on: push: branches: - master + - dev release: types: [published] From 475ffb1128f857b308252cd903b27222b9088dc8 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 3 Jun 2020 10:22:09 +0100 Subject: [PATCH 189/226] Remove dev test --- .github/workflows/awstest.yml | 1 - 1 file changed, 1 deletion(-) diff --git a/.github/workflows/awstest.yml b/.github/workflows/awstest.yml index c315d213..af704a87 100644 --- a/.github/workflows/awstest.yml +++ b/.github/workflows/awstest.yml @@ -6,7 +6,6 @@ on: push: branches: - master - - dev release: types: [published] From 0835128f4c28d6494fbdbb58df304a5528cd0b8e Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 3 Jun 2020 10:22:40 +0100 Subject: [PATCH 190/226] Use hg38 --- conf/test_full.config | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/conf/test_full.config b/conf/test_full.config index 94cd8e5e..e380e9c9 100755 --- a/conf/test_full.config +++ b/conf/test_full.config @@ -13,7 +13,7 @@ params { // Input data input = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/design_full.csv' - + // Genome references - genome = 'hg19' + genome = 'hg38' } From a3c459be79b712d2541fd853cc7607261f82a265 Mon Sep 17 00:00:00 2001 From: ggabernet Date: Tue, 16 Jun 2020 11:56:49 +0200 Subject: [PATCH 191/226] update awstest workflows --- .github/workflows/awsfulltest.yml | 36 +++++++++++++++ .github/workflows/awstest.yml | 75 ++++++++++++++++--------------- 2 files changed, 74 insertions(+), 37 deletions(-) create mode 100644 .github/workflows/awsfulltest.yml diff --git a/.github/workflows/awsfulltest.yml b/.github/workflows/awsfulltest.yml new file mode 100644 index 00000000..82098edf --- /dev/null +++ b/.github/workflows/awsfulltest.yml @@ -0,0 +1,36 @@ +name: nf-core AWS full size tests +# This workflow is triggered on releases. +# It runs the -profile 'test_full' on AWS batch + +on: + release: + types: [published] + +jobs: + run-awstest: + if: github.repository == 'nf-core/atacseq' + name: Run AWS test + runs-on: ubuntu-latest + steps: + - name: Setup Miniconda + uses: goanpeca/setup-miniconda@v1.0.2 + with: + auto-update-conda: true + python-version: 3.7 + - name: Install awscli + run: conda install -c conda-forge awscli + - name: Start AWS batch job + env: + AWS_ACCESS_KEY_ID: ${{secrets.AWSTEST_KEY_ID}} + AWS_SECRET_ACCESS_KEY: ${{secrets.AWSTEST_KEY_SECRET}} + TOWER_ACCESS_TOKEN: ${{secrets.AWSTEST_TOWER_TOKEN}} + #AWS_JOB_DEFINITION: ${{secrets.AWS_JOB_DEFINITION}} + AWS_JOB_QUEUE: ${{secrets.AWS_JOB_QUEUE}} + AWS_S3_BUCKET: ${{secrets.AWS_S3_BUCKET}} + run: | # Submits job to AWS batch using a 'nextflow-4GiB' job definition. Setting JVM options to "-XX:+UseG1GC" for more efficient garbage collection when staging remote files. + aws batch submit-job \ + --region eu-west-1 \ + --job-name nf-core-atacseq \ + --job-queue $AWS_JOB_QUEUE \ + --job-definition nextflow-4GiB \ + --container-overrides '{"command": ["nf-core/atacseq", "-r '"${GITHUB_SHA}"' -profile test_full --outdir s3://nf-core-awsmegatests/atacseq/results-'"${GITHUB_SHA}"' -w s3://nf-core-awsmegatests/atacseq/work-'"${GITHUB_SHA}"' -with-tower"], "environment": [{"name": "TOWER_ACCESS_TOKEN", "value": "'"$TOWER_ACCESS_TOKEN"'"}, {"name": "NXF_OPTS", "value": "-XX:+UseG1GC"}]}' diff --git a/.github/workflows/awstest.yml b/.github/workflows/awstest.yml index c315d213..c6923404 100644 --- a/.github/workflows/awstest.yml +++ b/.github/workflows/awstest.yml @@ -1,37 +1,38 @@ -name: nf-core AWS test -# This workflow is triggered on PRs to the master branch. -# It runs the -profile 'test_full' on AWS batch - -on: - push: - branches: - - master - - dev - release: - types: [published] - -jobs: - run-awstest: - if: github.repository == 'nf-core/atacseq' - name: Run AWS test - runs-on: ubuntu-latest - steps: - - name: Setup Miniconda - uses: goanpeca/setup-miniconda@v1.0.2 - with: - auto-update-conda: true - python-version: 3.7 - - name: Install awscli - run: conda install -c conda-forge awscli - - name: Start AWS batch job - env: - AWS_ACCESS_KEY_ID: ${{secrets.AWS_KEY_ID}} - AWS_SECRET_ACCESS_KEY: ${{secrets.AWS_KEY_SECRET}} - TOWER_ACCESS_TOKEN: ${{secrets.TOWER_ACCESS_TOKEN}} - run: | # Submits job to AWS batch using a 'nextflow-4GiB' instance. Setting JVM options to "-XX:+UseG1GC" for more efficient garbage collection when staging remote files. - aws batch submit-job \ - --region eu-west-1 \ - --job-name nf-core-atacseq \ - --job-queue 'default-8b3836e0-5eda-11ea-96e5-0a2c3f6a2a32' \ - --job-definition nextflow-4GiB \ - --container-overrides '{"command": ["nf-core/atacseq", "-r '"${GITHUB_SHA}"' -profile test_full --outdir s3://nf-core-awsmegatests/atacseq/results-'"${GITHUB_SHA}"' -w s3://nf-core-awsmegatests/atacseq/work-'"${GITHUB_SHA}"' -with-tower"], "environment": [{"name": "TOWER_ACCESS_TOKEN", "value": "'"$TOWER_ACCESS_TOKEN"'"}, {"name": "NXF_OPTS", "value": "-XX:+UseG1GC"}]}' +name: nf-core AWS test +# This workflow is triggered on push to master branch. +# It runs the -profile 'test' on AWS batch + +on: + push: + branches: + - master + - dev # just for testing purposes, to be removed + +jobs: + run-awstest: + if: github.repository == 'nf-core/atacseq' + name: Run AWS test + runs-on: ubuntu-latest + steps: + - name: Setup Miniconda + uses: goanpeca/setup-miniconda@v1.0.2 + with: + auto-update-conda: true + python-version: 3.7 + - name: Install awscli + run: conda install -c conda-forge awscli + - name: Start AWS batch job + env: + AWS_ACCESS_KEY_ID: ${{secrets.AWSTEST_KEY_ID}} + AWS_SECRET_ACCESS_KEY: ${{secrets.AWSTEST_KEY_SECRET}} + TOWER_ACCESS_TOKEN: ${{secrets.AWSTEST_TOWER_TOKEN}} + #AWS_JOB_DEFINITION: ${{secrets.AWS_JOB_DEFINITION}} + AWS_JOB_QUEUE: ${{secrets.AWS_JOB_QUEUE}} + AWS_S3_BUCKET: ${{secrets.AWS_S3_BUCKET}} + run: | # Submits job to AWS batch using a 'nextflow-4GiB' job definition. Setting JVM options to "-XX:+UseG1GC" for more efficient garbage collection when staging remote files. + aws batch submit-job \ + --region eu-west-1 \ + --job-name nf-core-atacseq \ + --job-queue $AWS_JOB_QUEUE \ + --job-definition nextflow-4GiB \ + --container-overrides '{"command": ["nf-core/atacseq", "-r '"${GITHUB_SHA}"' -profile test --outdir s3://nf-core-awsmegatests/atacseq/results-'"${GITHUB_SHA}"' -w s3://nf-core-awsmegatests/atacseq/work-'"${GITHUB_SHA}"' -with-tower"], "environment": [{"name": "TOWER_ACCESS_TOKEN", "value": "'"$TOWER_ACCESS_TOKEN"'"}, {"name": "NXF_OPTS", "value": "-XX:+UseG1GC"}]}' From 6c8cef753a1cbb004507003497460cdd2b112daf Mon Sep 17 00:00:00 2001 From: ggabernet Date: Tue, 16 Jun 2020 17:16:58 +0200 Subject: [PATCH 192/226] update awstest --- .github/workflows/awstest.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/.github/workflows/awstest.yml b/.github/workflows/awstest.yml index c6923404..56488beb 100644 --- a/.github/workflows/awstest.yml +++ b/.github/workflows/awstest.yml @@ -1,5 +1,5 @@ name: nf-core AWS test -# This workflow is triggered on push to master branch. +# This workflow is triggered on push to the master branch. # It runs the -profile 'test' on AWS batch on: From a21890e3a75ca4d9dd38ee492de610c35dab8071 Mon Sep 17 00:00:00 2001 From: ggabernet Date: Tue, 16 Jun 2020 17:22:18 +0200 Subject: [PATCH 193/226] update changelog --- CHANGELOG.md | 1 + 1 file changed, 1 insertion(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index 7d4aba66..1a13e46e 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -34,6 +34,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * [nf-core/chipseq#132](https://github.com/nf-core/chipseq/issues/132) - BigWig Error: sort: cannot create temporary file in '': Read-only file system * [nf-core/chipseq#154](https://github.com/nf-core/chipseq/issues/154) - computeMatrix.val.mat.gz files not zipped * Make executables in `bin/` compatible with Python 3 +* [#110](https://github.com/nf-core/atacseq/pull/110) - updated AWS test GitHub actions ### `Dependencies` From 229537f76403d91597ea86234703fb7262145dc6 Mon Sep 17 00:00:00 2001 From: ggabernet Date: Tue, 16 Jun 2020 19:49:59 +0200 Subject: [PATCH 194/226] awstest s3 bucket var --- .github/workflows/awstest.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/.github/workflows/awstest.yml b/.github/workflows/awstest.yml index 56488beb..0bf1ae3c 100644 --- a/.github/workflows/awstest.yml +++ b/.github/workflows/awstest.yml @@ -35,4 +35,4 @@ jobs: --job-name nf-core-atacseq \ --job-queue $AWS_JOB_QUEUE \ --job-definition nextflow-4GiB \ - --container-overrides '{"command": ["nf-core/atacseq", "-r '"${GITHUB_SHA}"' -profile test --outdir s3://nf-core-awsmegatests/atacseq/results-'"${GITHUB_SHA}"' -w s3://nf-core-awsmegatests/atacseq/work-'"${GITHUB_SHA}"' -with-tower"], "environment": [{"name": "TOWER_ACCESS_TOKEN", "value": "'"$TOWER_ACCESS_TOKEN"'"}, {"name": "NXF_OPTS", "value": "-XX:+UseG1GC"}]}' + --container-overrides '{"command": ["nf-core/atacseq", "-r '"${GITHUB_SHA}"' -profile test --outdir s3://'"${AWS_S3_BUCKET}"'/atacseq/results-'"${GITHUB_SHA}"' -w s3://'"${AWS_S3_BUCKET}"'/atacseq/work-'"${GITHUB_SHA}"' -with-tower"], "environment": [{"name": "TOWER_ACCESS_TOKEN", "value": "'"$TOWER_ACCESS_TOKEN"'"}, {"name": "NXF_OPTS", "value": "-XX:+UseG1GC"}]}' From 074bb151321d6a5631400dda25488209974912ee Mon Sep 17 00:00:00 2001 From: ggabernet Date: Wed, 17 Jun 2020 23:01:14 +0200 Subject: [PATCH 195/226] aws test update secrets names --- .github/workflows/awsfulltest.yml | 8 ++++---- .github/workflows/awstest.yml | 6 +++--- 2 files changed, 7 insertions(+), 7 deletions(-) diff --git a/.github/workflows/awsfulltest.yml b/.github/workflows/awsfulltest.yml index 82098edf..15f940dd 100644 --- a/.github/workflows/awsfulltest.yml +++ b/.github/workflows/awsfulltest.yml @@ -21,9 +21,9 @@ jobs: run: conda install -c conda-forge awscli - name: Start AWS batch job env: - AWS_ACCESS_KEY_ID: ${{secrets.AWSTEST_KEY_ID}} - AWS_SECRET_ACCESS_KEY: ${{secrets.AWSTEST_KEY_SECRET}} - TOWER_ACCESS_TOKEN: ${{secrets.AWSTEST_TOWER_TOKEN}} + AWS_ACCESS_KEY_ID: ${{secrets.AWS_ACCESS_KEY_ID}} + AWS_SECRET_ACCESS_KEY: ${{secrets.AWS_SECRET_ACCESS_KEY}} + TOWER_ACCESS_TOKEN: ${{secrets.AWS_TOWER_TOKEN}} #AWS_JOB_DEFINITION: ${{secrets.AWS_JOB_DEFINITION}} AWS_JOB_QUEUE: ${{secrets.AWS_JOB_QUEUE}} AWS_S3_BUCKET: ${{secrets.AWS_S3_BUCKET}} @@ -33,4 +33,4 @@ jobs: --job-name nf-core-atacseq \ --job-queue $AWS_JOB_QUEUE \ --job-definition nextflow-4GiB \ - --container-overrides '{"command": ["nf-core/atacseq", "-r '"${GITHUB_SHA}"' -profile test_full --outdir s3://nf-core-awsmegatests/atacseq/results-'"${GITHUB_SHA}"' -w s3://nf-core-awsmegatests/atacseq/work-'"${GITHUB_SHA}"' -with-tower"], "environment": [{"name": "TOWER_ACCESS_TOKEN", "value": "'"$TOWER_ACCESS_TOKEN"'"}, {"name": "NXF_OPTS", "value": "-XX:+UseG1GC"}]}' + --container-overrides '{"command": ["nf-core/atacseq", "-r '"${GITHUB_SHA}"' -profile test_full --outdir s3://'"${AWS_S3_BUCKET}"'/atacseq/results-'"${GITHUB_SHA}"' -w s3://'"${AWS_S3_BUCKET}"'/atacseq/work-'"${GITHUB_SHA}"' -with-tower"], "environment": [{"name": "TOWER_ACCESS_TOKEN", "value": "'"$TOWER_ACCESS_TOKEN"'"}, {"name": "NXF_OPTS", "value": "-XX:+UseG1GC"}]}' diff --git a/.github/workflows/awstest.yml b/.github/workflows/awstest.yml index 0bf1ae3c..fae13576 100644 --- a/.github/workflows/awstest.yml +++ b/.github/workflows/awstest.yml @@ -23,9 +23,9 @@ jobs: run: conda install -c conda-forge awscli - name: Start AWS batch job env: - AWS_ACCESS_KEY_ID: ${{secrets.AWSTEST_KEY_ID}} - AWS_SECRET_ACCESS_KEY: ${{secrets.AWSTEST_KEY_SECRET}} - TOWER_ACCESS_TOKEN: ${{secrets.AWSTEST_TOWER_TOKEN}} + AWS_ACCESS_KEY_ID: ${{secrets.AWS_ACCESS_KEY_ID}} + AWS_SECRET_ACCESS_KEY: ${{secrets.AWS_SECRET_ACCESS_KEY}} + TOWER_ACCESS_TOKEN: ${{secrets.AWS_TOWER_TOKEN}} #AWS_JOB_DEFINITION: ${{secrets.AWS_JOB_DEFINITION}} AWS_JOB_QUEUE: ${{secrets.AWS_JOB_QUEUE}} AWS_S3_BUCKET: ${{secrets.AWS_S3_BUCKET}} From 04c3dd53ba4000c682c1b2ef77676ac6dc5f357d Mon Sep 17 00:00:00 2001 From: Phil Ewels Date: Wed, 24 Jun 2020 08:24:18 +0200 Subject: [PATCH 196/226] Remove required group --- nextflow_schema.json | 3 +-- 1 file changed, 1 insertion(+), 2 deletions(-) diff --git a/nextflow_schema.json b/nextflow_schema.json index 83c5b73e..c9174afd 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -535,6 +535,5 @@ }, "fa_icon": "fas fa-file-import" } - }, - "required": ["Input/output options"] + } } From 27fd7b1b2287247ad30d868cabf4919a582decda Mon Sep 17 00:00:00 2001 From: Phil Ewels Date: Thu, 25 Jun 2020 11:03:05 +0200 Subject: [PATCH 197/226] Remove default value from schema for required input --- nextflow_schema.json | 1 - 1 file changed, 1 deletion(-) diff --git a/nextflow_schema.json b/nextflow_schema.json index c9174afd..b8f88985 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -10,7 +10,6 @@ "properties": { "input": { "type": "string", - "default": "./design.csv", "description": "Comma-separated file containing information about the samples in the experiment.", "help_text": "You will need to create a design file with information about the samples in your experiment before running the pipeline. Use this parameter to specify its location. It has to be a comma-separated file with 4 columns, and a header row. See [usage docs](https://nf-co.re/atacseq/docs/usage#--input).", "fa_icon": "fas fa-file-csv" From 379e4841d5650ed1d3789dd35a371df3d4883a6c Mon Sep 17 00:00:00 2001 From: Phil Ewels Date: Sat, 27 Jun 2020 01:05:12 +0200 Subject: [PATCH 198/226] Added additional parameters to the JSON schema --- nextflow_schema.json | 23 +++++++++++++++++++---- 1 file changed, 19 insertions(+), 4 deletions(-) diff --git a/nextflow_schema.json b/nextflow_schema.json index b8f88985..5d739279 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -10,7 +10,7 @@ "properties": { "input": { "type": "string", - "description": "Comma-separated file containing information about the samples in the experiment.", + "description": "Path to comma-separated file containing information about the samples in the experiment.", "help_text": "You will need to create a design file with information about the samples in your experiment before running the pipeline. Use this parameter to specify its location. It has to be a comma-separated file with 4 columns, and a header row. See [usage docs](https://nf-co.re/atacseq/docs/usage#--input).", "fa_icon": "fas fa-file-csv" }, @@ -35,7 +35,7 @@ }, "outdir": { "type": "string", - "description": "The output directory where the results will be saved.", + "description": "Path to the output directory where the results will be saved.", "default": "./results", "fa_icon": "fas fa-folder-open", "help_text": "" @@ -76,7 +76,7 @@ }, "bwa_index": { "type": "string", - "description": "Full path to directory containing BWA index including base name i.e. /path/to/index/genome.fa.", + "description": "Full path to directory containing BWA index including base name. i.e. `/path/to/index/genome.fa.`", "fa_icon": "fas fa-bezier-curve", "help_text": "" }, @@ -279,6 +279,13 @@ "description": "Skip consensus peak generation, annotation and counting.", "fa_icon": "fas fa-forward", "help_text": "" + }, + "anno_readme": { + "type": "string", + "description": "Path to annotation readme file from AWS iGenomes", + "hidden": true, + "help_text": "This parameter is typically set within institutional iGenomes config files.", + "fa_icon": "fas fa-file-alt" } }, "fa_icon": "fas fa-chart-area" @@ -348,6 +355,14 @@ "description": "Skip IGV.", "fa_icon": "fas fa-forward" }, + "skip_peak_qc": { + "type": "string", + "fa_icon": "fas fa-forward" + }, + "skip_peak_annotation": { + "type": "string", + "fa_icon": "fas fa-forward" + }, "skip_multiqc": { "type": "boolean", "default": false, @@ -535,4 +550,4 @@ "fa_icon": "fas fa-file-import" } } -} +} \ No newline at end of file From fb4f4d973d1abf2769d09cb2d9ba9751254ebe55 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 30 Jun 2020 11:23:00 +0100 Subject: [PATCH 199/226] Schema edits --- nextflow_schema.json | 79 +++++++++++++++++++++----------------------- 1 file changed, 38 insertions(+), 41 deletions(-) diff --git a/nextflow_schema.json b/nextflow_schema.json index 5d739279..0a570a76 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -25,7 +25,7 @@ "type": "integer", "default": 0, "description": "Estimated fragment size used to extend single-end reads.", - "fa_icon": "fas fa-align-justify", + "fa_icon": "fas fa-chart-area", "help_text": "" }, "seq_center": { @@ -76,7 +76,7 @@ }, "bwa_index": { "type": "string", - "description": "Full path to directory containing BWA index including base name. i.e. `/path/to/index/genome.fa.`", + "description": "Full path to directory containing BWA index including base name. i.e. `/path/to/index/genome.fa`.", "fa_icon": "fas fa-bezier-curve", "help_text": "" }, @@ -102,7 +102,7 @@ "type": "string", "description": "Path to blacklist regions in BED format, used for filtering alignments.", "help_text": "If provided, alignments that overlap with the regions in this file will be filtered out (see [ENCODE blacklists](https://sites.google.com/site/anshulkundaje/projects/blacklists)). The file should be in BED format. Blacklisted regions for *GRCh37*, *GRCh38*, *GRCm38*, *hg19*, *hg38*, *mm10* are bundled with the pipeline in the [`blacklists`](../assets/blacklists/) directory, and as such will be automatically used if any of those genomes are specified with the `--genome` parameter.", - "fa_icon": "fas fa-clipboard-list" + "fa_icon": "fas fa-book-dead" }, "mito_name": { "type": "string", @@ -273,19 +273,24 @@ "fa_icon": "fas fa-save", "help_text": "" }, + "skip_peak_qc": { + "type": "boolean", + "fa_icon": "fas fa-forward", + "description": "Skip MACS2 peak QC plot generation.", + "default": false + }, + "skip_peak_annotation": { + "type": "boolean", + "fa_icon": "fas fa-forward", + "description": "Skip annotation of MACS2 and consensus peaks with HOMER.", + "default": false + }, "skip_consensus_peaks": { "type": "boolean", "default": false, "description": "Skip consensus peak generation, annotation and counting.", "fa_icon": "fas fa-forward", "help_text": "" - }, - "anno_readme": { - "type": "string", - "description": "Path to annotation readme file from AWS iGenomes", - "hidden": true, - "help_text": "This parameter is typically set within institutional iGenomes config files.", - "fa_icon": "fas fa-file-alt" } }, "fa_icon": "fas fa-chart-area" @@ -355,14 +360,6 @@ "description": "Skip IGV.", "fa_icon": "fas fa-forward" }, - "skip_peak_qc": { - "type": "string", - "fa_icon": "fas fa-forward" - }, - "skip_peak_annotation": { - "type": "string", - "fa_icon": "fas fa-forward" - }, "skip_multiqc": { "type": "boolean", "default": false, @@ -458,21 +455,20 @@ "help": { "type": "boolean", "description": "Display help text.", - "hidden": true, "fa_icon": "fas fa-question-circle", - "default": false + "default": false, + "hidden": true }, "fingerprint_bins": { "type": "integer", "default": 500000, - "hidden": true, "description": "Number of genomic bins to use when calculating deepTools fingerprint plot.", - "fa_icon": "fas fa-dumpster" + "fa_icon": "fas fa-dumpster", + "hidden": true }, "publish_dir_mode": { "type": "string", "default": "copy", - "hidden": true, "description": "Method used to save pipeline results to output directory.", "help_text": "The Nextflow `publishDir` option specifies which intermediate files should be saved to the output directory. This option tells the pipeline what method should be used to move these files. See [Nextflow docs](https://www.nextflow.io/docs/latest/process.html#publishdir) for details.", "fa_icon": "fas fa-copy", @@ -483,68 +479,69 @@ "copy", "copyNoFollow", "mov" - ] + ], + "hidden": true }, "name": { "type": "string", "description": "Workflow name.", - "fa_icon": "fas fa-fingerprint", - "hidden": true, - "help_text": "A custom name for the pipeline run. Unlike the core nextflow `-name` option with one hyphen this parameter can be reused multiple times, for example if using `-resume`. Passed through to steps such as MultiQC and used for things like report filenames and titles." + "fa_icon": "fas fa-address-card", + "help_text": "A custom name for the pipeline run. Unlike the core nextflow `-name` option with one hyphen this parameter can be reused multiple times, for example if using `-resume`. Passed through to steps such as MultiQC and used for things like report filenames and titles.", + "hidden": true }, "email_on_fail": { "type": "string", "description": "Email address for completion summary, only when pipeline fails.", "fa_icon": "fas fa-exclamation-triangle", "pattern": "^([a-zA-Z0-9_\\-\\.]+)@([a-zA-Z0-9_\\-\\.]+)\\.([a-zA-Z]{2,5})$", - "hidden": true, - "help_text": "An email address to send a summary email to when the pipeline is completed - ONLY sent if the pipeline does not exit successfully." + "help_text": "An email address to send a summary email to when the pipeline is completed - ONLY sent if the pipeline does not exit successfully.", + "hidden": true }, "plaintext_email": { "type": "boolean", "description": "Send plain-text email instead of HTML.", "fa_icon": "fas fa-remove-format", - "hidden": true, "default": false, - "help_text": "" + "help_text": "", + "hidden": true }, "max_multiqc_email_size": { "type": "string", "description": "File size limit when attaching MultiQC reports to summary emails.", "default": "25.MB", "fa_icon": "fas fa-file-upload", - "hidden": true, - "help_text": "" + "help_text": "", + "hidden": true }, "monochrome_logs": { "type": "boolean", "description": "Do not use coloured log outputs.", "fa_icon": "fas fa-palette", - "hidden": true, "default": false, - "help_text": "" + "help_text": "", + "hidden": true }, "multiqc_config": { "type": "string", "description": "Custom config file to supply to MultiQC.", "fa_icon": "fas fa-cog", - "hidden": true, - "help_text": "" + "help_text": "", + "hidden": true }, "tracedir": { "type": "string", "description": "Directory to keep pipeline Nextflow logs and reports.", "default": "${params.outdir}/pipeline_info", "fa_icon": "fas fa-cogs", - "hidden": true, - "help_text": "" + "help_text": "", + "hidden": true }, "clusterOptions": { "type": "string", "description": "Arguments passed to Nextflow clusterOptions.", - "hidden": true, "fa_icon": "fas fa-network-wired", - "help_text": "" + "help_text": "", + "hidden": true } }, "fa_icon": "fas fa-file-import" From 379786646dea72d2e09119a9b0cecf6992cdc6c9 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 30 Jun 2020 13:08:21 +0100 Subject: [PATCH 200/226] Update to latest on tools:dev --- .github/workflows/ci.yml | 105 +++++++++++++++++++++++++++++++++------ 1 file changed, 90 insertions(+), 15 deletions(-) mode change 100644 => 100755 .github/workflows/ci.yml diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml old mode 100644 new mode 100755 index 86b5670e..675965f5 --- a/.github/workflows/ci.yml +++ b/.github/workflows/ci.yml @@ -1,50 +1,125 @@ name: nf-core CI -# This workflow is triggered on pushes and PRs to the repository. +# This workflow is triggered on releases and pull-requests. # It runs the pipeline with the minimal test dataset to check that it completes without any syntax errors -on: [push, pull_request] +on: + push: + branches: + - dev + pull_request: + release: + types: [published] jobs: test: + name: Run workflow tests + # Only run on push if this is the nf-core dev branch (merged PRs) + if: ${{ github.event_name != 'push' || (github.event_name == 'push' && github.repository == 'nf-core/atacseq') }} + runs-on: ubuntu-latest env: NXF_VER: ${{ matrix.nxf_ver }} NXF_ANSI_LOG: false - runs-on: ubuntu-latest strategy: matrix: # Nextflow versions: check pipeline minimum and current latest nxf_ver: ['19.10.0', ''] steps: - - uses: actions/checkout@v2 - - name: Install Nextflow - run: | - wget -qO- get.nextflow.io | bash - sudo mv nextflow /usr/local/bin/ + - name: Check out pipeline code + uses: actions/checkout@v2 + + - name: Check if Dockerfile or Conda environment changed + uses: technote-space/get-diff-action@v1 + with: + PREFIX_FILTER: | + Dockerfile + environment.yml + + - name: Build new docker image + if: env.GIT_DIFF + run: docker build --no-cache . -t nfcore/atacseq:dev + - name: Pull docker image + if: ${{ !env.GIT_DIFF }} run: | docker pull nfcore/atacseq:dev docker tag nfcore/atacseq:dev nfcore/atacseq:dev + + - name: Install Nextflow + run: | + wget -qO- get.nextflow.io | bash + sudo mv nextflow /usr/local/bin/ + - name: Run pipeline with test data run: | nextflow run ${GITHUB_WORKSPACE} -profile test,docker parameters: + name: Test workflow parameters + if: ${{ github.event_name != 'push' || (github.event_name == 'push' && github.repository == 'nf-core/atacseq') }} + runs-on: ubuntu-latest env: NXF_VER: '19.10.0' NXF_ANSI_LOG: false - runs-on: ubuntu-latest strategy: matrix: parameters: [--single_end, --skip_trimming, --skip_merge_replicates, --skip_consensus_peaks] steps: - - uses: actions/checkout@v2 - - name: Install Nextflow - run: | - wget -qO- get.nextflow.io | bash - sudo mv nextflow /usr/local/bin/ + - name: Check out pipeline code + uses: actions/checkout@v2 + + - name: Check if Dockerfile or Conda environment changed + uses: technote-space/get-diff-action@v1 + with: + PREFIX_FILTER: | + Dockerfile + environment.yml + + - name: Build new docker image + if: env.GIT_DIFF + run: docker build --no-cache . -t nfcore/atacseq:dev + - name: Pull docker image + if: ${{ !env.GIT_DIFF }} run: | docker pull nfcore/atacseq:dev docker tag nfcore/atacseq:dev nfcore/atacseq:dev - - name: Run pipeline with various options + + - name: Install Nextflow + run: | + wget -qO- get.nextflow.io | bash + sudo mv nextflow /usr/local/bin/ + + - name: Run pipeline with various parameters run: | nextflow run ${GITHUB_WORKSPACE} -profile test,docker ${{ matrix.parameters }} + + push_dockerhub: + name: Push new Docker image to Docker Hub + runs-on: ubuntu-latest + # Only run if the tests passed + needs: test + # Only run for the nf-core repo, for releases and merged PRs + if: ${{ github.repository == 'nf-core/atacseq' && (github.event_name == 'release' || github.event_name == 'push') }} + env: + DOCKERHUB_USERNAME: ${{ secrets.DOCKERHUB_USERNAME }} + DOCKERHUB_PASS: ${{ secrets.DOCKERHUB_PASS }} + steps: + - name: Check out pipeline code + uses: actions/checkout@v2 + + - name: Build new docker image + run: docker build --no-cache . -t nfcore/atacseq:latest + + - name: Push Docker image to DockerHub (dev) + if: ${{ github.event_name == 'push' }} + run: | + echo "$DOCKERHUB_PASS" | docker login -u "$DOCKERHUB_USERNAME" --password-stdin + docker tag nfcore/atacseq:latest nfcore/atacseq:dev + docker push nfcore/atacseq:dev + + - name: Push Docker image to DockerHub (release) + if: ${{ github.event_name == 'release' }} + run: | + echo "$DOCKERHUB_PASS" | docker login -u "$DOCKERHUB_USERNAME" --password-stdin + docker push nfcore/atacseq:latest + docker tag nfcore/atacseq:latest nfcore/atacseq:${{ github.event.release.tag_name }} + docker push nfcore/atacseq:${{ github.event.release.tag_name }} From aacb29ebf8dbf436aab869f67f3c9dae61413dea Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 30 Jun 2020 13:08:29 +0100 Subject: [PATCH 201/226] Update to latest on tools:dev --- .github/workflows/linting.yml | 13 ++++++++++++- 1 file changed, 12 insertions(+), 1 deletion(-) mode change 100644 => 100755 .github/workflows/linting.yml diff --git a/.github/workflows/linting.yml b/.github/workflows/linting.yml old mode 100644 new mode 100755 index 1e0827a8..eb66c144 --- a/.github/workflows/linting.yml +++ b/.github/workflows/linting.yml @@ -33,18 +33,29 @@ jobs: nf-core: runs-on: ubuntu-latest steps: - - uses: actions/checkout@v2 + + - name: Check out pipeline code + uses: actions/checkout@v2 + - name: Install Nextflow run: | wget -qO- get.nextflow.io | bash sudo mv nextflow /usr/local/bin/ + - uses: actions/setup-python@v1 with: python-version: '3.6' architecture: 'x64' + - name: Install dependencies run: | python -m pip install --upgrade pip pip install nf-core + - name: Run nf-core lint + env: + GITHUB_COMMENTS_URL: ${{ github.event.pull_request.comments_url }} + GITHUB_TOKEN: ${{ secrets.GITHUB_TOKEN }} + GITHUB_PR_COMMIT: ${{ github.event.pull_request.head.sha }} run: nf-core lint ${GITHUB_WORKSPACE} + From 06cc09236696251a0d87245ea79b6b0c5c402777 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 30 Jun 2020 13:23:36 +0100 Subject: [PATCH 202/226] Add Slack badge --- README.md | 35 +++++++++++++++++------------------ 1 file changed, 17 insertions(+), 18 deletions(-) diff --git a/README.md b/README.md index d9ae7a95..28121d11 100644 --- a/README.md +++ b/README.md @@ -3,10 +3,11 @@ [![GitHub Actions CI Status](https://github.com/nf-core/atacseq/workflows/nf-core%20CI/badge.svg)](https://github.com/nf-core/atacseq/actions) [![GitHub Actions Linting Status](https://github.com/nf-core/atacseq/workflows/nf-core%20linting/badge.svg)](https://github.com/nf-core/atacseq/actions) [![Nextflow](https://img.shields.io/badge/nextflow-%E2%89%A519.10.0-brightgreen.svg)](https://www.nextflow.io/) +[![DOI](https://zenodo.org/badge/DOI/10.5281/zenodo.2634132.svg)](https://doi.org/10.5281/zenodo.2634132) [![install with bioconda](https://img.shields.io/badge/install%20with-bioconda-brightgreen.svg)](http://bioconda.github.io/) [![Docker](https://img.shields.io/docker/automated/nfcore/atacseq.svg)](https://hub.docker.com/r/nfcore/atacseq) -[![DOI](https://zenodo.org/badge/DOI/10.5281/zenodo.2634132.svg)](https://doi.org/10.5281/zenodo.2634132) +![[Get help on Slack](http://img.shields.io/badge/slack-nf--core%20%23atacseq-4A154B?logo=slack)](https://nfcore.slack.com/channels/atacseq) ## Introduction @@ -59,23 +60,23 @@ The pipeline is built using [Nextflow](https://www.nextflow.io), a workflow tool ## Quick Start -i. Install [`nextflow`](https://nf-co.re/usage/installation) +1. Install [`nextflow`](https://nf-co.re/usage/installation) -ii. Install either [`Docker`](https://docs.docker.com/engine/installation/) or [`Singularity`](https://www.sylabs.io/guides/3.0/user-guide/) for full pipeline reproducibility (please only use [`Conda`](https://conda.io/miniconda.html) as a last resort; see [docs](https://nf-co.re/usage/configuration#basic-configuration-profiles)) +2. Install either [`Docker`](https://docs.docker.com/engine/installation/) or [`Singularity`](https://www.sylabs.io/guides/3.0/user-guide/) for full pipeline reproducibility _(please only use [`Conda`](https://conda.io/miniconda.html) as a last resort; see [docs](https://nf-co.re/usage/configuration#basic-configuration-profiles))_ -iii. Download the pipeline and test it on a minimal dataset with a single command +3. Download the pipeline and test it on a minimal dataset with a single command: -```bash -nextflow run nf-core/atacseq -profile test, -``` + ```bash + nextflow run nf-core/atacseq -profile test, + ``` -> Please check [nf-core/configs](https://github.com/nf-core/configs#documentation) to see if a custom config file to run nf-core pipelines already exists for your Institute. If so, you can simply use `-profile ` in your command. This will enable either `docker` or `singularity` and set the appropriate execution settings for your local compute environment. + > Please check [nf-core/configs](https://github.com/nf-core/configs#documentation) to see if a custom config file to run nf-core pipelines already exists for your Institute. If so, you can simply use `-profile ` in your command. This will enable either `docker` or `singularity` and set the appropriate execution settings for your local compute environment. -iv. Start running your own analysis! +4. Start running your own analysis! -```bash -nextflow run nf-core/atacseq -profile --input design.csv --genome GRCh37 -``` + ```bash + nextflow run nf-core/atacseq -profile --input design.csv --genome GRCh37 + ``` See [usage docs](docs/usage.md) for all of the available options when running the pipeline. @@ -98,20 +99,20 @@ The pipeline was originally written by [The Bioinformatics & Biostatistics Group The pipeline was developed by [Harshil Patel](mailto:harshil.patel@crick.ac.uk). -The [nf-core/rnaseq](https://github.com/nf-core/rnaseq) and [nf-core/chipseq](https://github.com/nf-core/chipseq) pipelines developed by Phil Ewels were initially used as a template for this pipeline. Many thanks to Phil for all of his help and advice, and the team at SciLifeLab. - -Many thanks to others who have helped out and contributed along the way too, including (but not limited to): [@apeltzer](https://github.com/apeltzer), [@crickbabs](https://github.com/crickbabs), [drewjbeh](https://github.com/drewjbeh), [@houghtos](https://github.com/houghtos), [@jinmingda](https://github.com/jinmingda), [@ktrns](https://github.com/ktrns), [@MaxUlysse](https://github.com/MaxUlysse), [@mashehu](https://github.com/mashehu), [@micans](https://github.com/micans), [@pditommaso](https://github.com/pditommaso) and [@sven1103](https://github.com/sven1103). +Many thanks to others who have helped out and contributed along the way too, including (but not limited to): [@ewels](https://github.com/ewels), [@apeltzer](https://github.com/apeltzer), [@crickbabs](https://github.com/crickbabs), [drewjbeh](https://github.com/drewjbeh), [@houghtos](https://github.com/houghtos), [@jinmingda](https://github.com/jinmingda), [@ktrns](https://github.com/ktrns), [@MaxUlysse](https://github.com/MaxUlysse), [@mashehu](https://github.com/mashehu), [@micans](https://github.com/micans), [@pditommaso](https://github.com/pditommaso) and [@sven1103](https://github.com/sven1103). ## Contributions and Support If you would like to contribute to this pipeline, please see the [contributing guidelines](.github/CONTRIBUTING.md). -For further information or help, don't hesitate to get in touch on [Slack](https://nfcore.slack.com/channels/atacseq) (you can join with [this invite](https://nf-co.re/join/slack)). +For further information or help, don't hesitate to get in touch on the [Slack `#atacseq` channel](https://nfcore.slack.com/channels/atacseq) (you can join with [this invite](https://nf-co.re/join/slack)). ## Citation If you use nf-core/atacseq for your analysis, please cite it using the following doi: [10.5281/zenodo.2634132](https://doi.org/10.5281/zenodo.2634132) +An extensive list of references for the tools used by the pipeline can be found in the [`CITATIONS.md`](CITATIONS.md) file. + You can cite the `nf-core` publication as follows: > **The nf-core framework for community-curated bioinformatics pipelines.** @@ -120,5 +121,3 @@ You can cite the `nf-core` publication as follows: > > _Nat Biotechnol._ 2020 Feb 13. doi: [10.1038/s41587-020-0439-x](https://dx.doi.org/10.1038/s41587-020-0439-x). > ReadCube: [Full Access Link](https://rdcu.be/b1GjZ) - -An extensive list of references for the tools used by the pipeline can be found in the [`CITATIONS.md`](CITATIONS.md) file. From b326f5be69b201962ef3c3cd0df63cc738dadc9e Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 30 Jun 2020 14:42:10 +0100 Subject: [PATCH 203/226] Relax linting --- .github/markdownlint.yml | 14 +++++++++----- 1 file changed, 9 insertions(+), 5 deletions(-) diff --git a/.github/markdownlint.yml b/.github/markdownlint.yml index 96b12a70..9aae63a0 100644 --- a/.github/markdownlint.yml +++ b/.github/markdownlint.yml @@ -1,5 +1,9 @@ -# Markdownlint configuration file -default: true, -line-length: false -no-duplicate-header: - siblings_only: true +# Markdownlint configuration file +default: true, +line-length: false +no-duplicate-header: + siblings_only: true +MD033: + allowed_elements: [details, summary] +MD007: + indent: 4 From 647a093440db5d5074052ededd1efa048c15b57c Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 30 Jun 2020 14:42:19 +0100 Subject: [PATCH 204/226] Reorganise docs --- docs/output.md | 418 ++++++++++++++++++++++++------------------------- 1 file changed, 206 insertions(+), 212 deletions(-) diff --git a/docs/output.md b/docs/output.md index 417be661..db87dab3 100755 --- a/docs/output.md +++ b/docs/output.md @@ -1,4 +1,4 @@ -# nf-core/atacseq: Output +# ![nf-core/atacseq](images/nf-core-atacseq_logo.png) This document describes the output produced by the pipeline. Most of the plots are taken from the MultiQC report, which summarises results at the end of the pipeline. @@ -14,326 +14,320 @@ The directories listed below will be created in the output directory after the p The initial QC and alignments are performed at the library-level e.g. if the same library has been sequenced more than once to increase sequencing depth. This has the advantage of being able to assess each library individually, and the ability to process multiple libraries from the same sample in parallel. -1. **Raw read QC** +### Raw read QC - *Documentation*: - [FastQC](http://www.bioinformatics.babraham.ac.uk/projects/fastqc/Help/) +[FastQC](http://www.bioinformatics.babraham.ac.uk/projects/fastqc/Help/) gives general quality metrics about your reads. It provides information about the quality score distribution across your reads, the per base sequence content (%A/C/G/T). You get information about adapter contamination and other overrepresented sequences. - *Description*: - FastQC gives general quality metrics about your reads. It provides information about the quality score distribution across your reads, the per base sequence content (%A/C/G/T). You get information about adapter contamination and other overrepresented sequences. +
+ Output files - *Output directories*: - * `fastqc/` - FastQC `*.html` files for read 1 (*and read2 if paired-end*) **before** adapter trimming. - * `fastqc/zips/` - FastQC `*.zip` files for read 1 (*and read2 if paired-end*) **before** adapter trimming. +* `fastqc/` +FastQC `*.html` files for read 1 (*and read2 if paired-end*) **before** adapter trimming. +* `fastqc/zips/` +FastQC `*.zip` files for read 1 (*and read2 if paired-end*) **before** adapter trimming. -2. **Adapter trimming** +
- *Documentation*: - [Trim Galore!](https://www.bioinformatics.babraham.ac.uk/projects/trim_galore/) +### Adapter trimming - *Description*: - Trim Galore! is a wrapper tool around Cutadapt and FastQC to consistently apply quality and adapter trimming to FastQ files. By default, Trim Galore! will automatically detect and trim the appropriate adapter sequence. For most ATAC-seq datasets this will be the Nextera adapter sequence 'CTGTCTCTTATA'. See [`usage.md`](usage.md) for more details about the trimming options. +[Trim Galore!](https://www.bioinformatics.babraham.ac.uk/projects/trim_galore/) is a wrapper tool around Cutadapt and FastQC to consistently apply quality and adapter trimming to FastQ files. By default, Trim Galore! will automatically detect and trim the appropriate adapter sequence. For most ATAC-seq datasets this will be the Nextera adapter sequence 'CTGTCTCTTATA'. See [`usage.md`](usage.md) for more details about the trimming options. - ![MultiQC - Cutadapt trimmed sequence plot](images/mqc_cutadapt_plot.png) +
+ Output files - *Output directories*: - * `trim_galore/` - If `--save_trimmed` is specified FastQ files **after** adapter trimming will be placed in this directory. - * `trim_galore/logs/` - `*.log` files generated by Trim Galore!. - * `trim_galore/fastqc/` - FastQC `*.html` files for read 1 (*and read2 if paired-end*) **after** adapter trimming. - * `trim_galore/fastqc/zips/` - FastQC `*.zip` files for read 1 (*and read2 if paired-end*) **after** adapter trimming. +* `trim_galore/` + If `--save_trimmed` is specified FastQ files **after** adapter trimming will be placed in this directory. +* `trim_galore/logs/` + `*.log` files generated by Trim Galore!. +* `trim_galore/fastqc/` + FastQC `*.html` files for read 1 (*and read2 if paired-end*) **after** adapter trimming. +* `trim_galore/fastqc/zips/` + FastQC `*.zip` files for read 1 (*and read2 if paired-end*) **after** adapter trimming. -3. **Alignment** +
- *Documentation*: - [BWA](http://bio-bwa.sourceforge.net/bwa.shtml), [SAMtools](http://samtools.sourceforge.net/) +![MultiQC - Cutadapt trimmed sequence plot](images/mqc_cutadapt_plot.png) - *Description*: - Adapter-trimmed reads are mapped to the reference assembly using BWA. A genome index is required to run BWA so if this is not provided explicitly using the `--bwa_index` parameter then it will be created automatically from the genome fasta input. The index creation process can take a while for larger genomes so it is possible to use the `--save_reference` parameter to save the indices for future pipeline runs, reducing processing times. +### Alignment - ![MultiQC - SAMtools stats plot](images/mqc_samtools_stats_plot.png) +Adapter-trimmed reads are mapped to the reference assembly using [BWA](http://bio-bwa.sourceforge.net/bwa.shtml). A genome index is required to run BWA so if this is not provided explicitly using the `--bwa_index` parameter then it will be created automatically from the genome fasta input. The index creation process can take a while for larger genomes so it is possible to use the `--save_reference` parameter to save the indices for future pipeline runs, reducing processing times. - ![MultiQC - SAMtools idxstats plot](images/mqc_samtools_idxstats_plot.png) +
+ Output files - File names in the resulting directory (i.e. `bwa/library/`) will have the '`.Lb.`' (**L**i**b**rary) suffix. +* `bwa/library/` + The files resulting from the alignment of individual libraries are not saved by default so this directory will not be present in your results. You can override this behaviour with the use of the `--save_align_intermeds` flag in which case it will contain the coordinate sorted alignment files in [`*.bam`](https://samtools.github.io/hts-specs/SAMv1.pdf) format. +* `bwa/library/samtools_stats/` + SAMtools `*.flagstat`, `*.idxstats` and `*.stats` files generated from the alignment files. - *Output directories*: - * `bwa/library/` - The files resulting from the alignment of individual libraries are not saved by default so this directory will not be present in your results. You can override this behaviour with the use of the `--save_align_intermeds` flag in which case it will contain the coordinate sorted alignment files in [`*.bam`](https://samtools.github.io/hts-specs/SAMv1.pdf) format. - * `bwa/library/samtools_stats/` - SAMtools `*.flagstat`, `*.idxstats` and `*.stats` files generated from the alignment files. +> **NB:** File names in the resulting directory (i.e. `bwa/library/`) will have the '`.Lb.`' suffix. + +
+ +![MultiQC - SAMtools stats plot](images/mqc_samtools_stats_plot.png) + +![MultiQC - SAMtools idxstats plot](images/mqc_samtools_idxstats_plot.png) ## Merged library-level analysis The library-level alignments associated with the same sample are merged and subsequently used for the downstream analyses. -1. **Alignment merging, duplicate marking, filtering and QC** +### Alignment merging, duplicate marking, filtering and QC + +[Picard MergeSamFiles and MarkDuplicates](https://broadinstitute.github.io/picard/command-line-overview.html) are used in combination to merge the alignments, and for the marking of duplicates, respectively. If you only have one library for any given replicate then the merging step isnt carried out because the library-level and merged library-level BAM files will be exactly the same. + +Read duplicate marking is carried out using the Picard MarkDuplicates command. Duplicate reads are generally removed from the aligned reads to mitigate for fragments in the library that may have been sequenced more than once due to PCR biases. There is an option to keep duplicate reads with the `--keep_dups` parameter but its generally recommended to remove them to avoid the wrong interpretation of the results. A similar option has been provided to keep reads that are multi-mapped - `--keep_multi_map`. Other steps have been incorporated into the pipeline to filter the resulting alignments - see [`main README.md`](../README.md) for a more comprehensive listing, and the tools used at each step. A selection of alignment-based QC metrics generated by Picard CollectMultipleMetrics and MarkDuplicates will be included in the MultiQC report. + +Certain cell types and tissues yield an enormous fraction (typically 20–80%) of unusable sequences of mitochondrial origin. This is a known problem that is specific to ATAC-seq library preps - see [Montefiori et al. 2017](https://www.nature.com/articles/s41598-017-02547-w). There is an option to keep these reads using the `--keep_mito` parameter but its generally recommended to remove these in order to get a more reliable assessment of the duplication rate from the rest of the genome, and to avoid any biases in the downstream analyses. - *Documentation*: - [picard](https://broadinstitute.github.io/picard/command-line-overview.html), [SAMtools](http://samtools.sourceforge.net/), [BEDTools](https://bedtools.readthedocs.io/en/latest/), [BAMTools](https://github.com/pezmaster31/bamtools/wiki/Tutorial_Toolkit_BamTools-1.0.pdf), [Pysam](https://pysam.readthedocs.io/en/latest/api.html), [Preseq](http://smithlabresearch.org/software/preseq/) +The [Preseq](http://smithlabresearch.org/software/preseq/) package is aimed at predicting and estimating the complexity of a genomic sequencing library, equivalent to predicting and estimating the number of redundant reads from a given sequencing depth and how many will be expected from additional sequencing using an initial sequencing experiment. The estimates can then be used to examine the utility of further sequencing, optimize the sequencing depth, or to screen multiple libraries to avoid low complexity samples. The dashed line shows a perfectly complex library where total reads = unique reads. Note that these are predictive numbers only, not absolute. The MultiQC plot can sometimes give extreme sequencing depth on the X axis - click and drag from the left side of the plot to zoom in on more realistic numbers. - *Description*: - Picard MergeSamFiles and MarkDuplicates are used in combination to merge the alignments, and for the marking of duplicates, respectively. If you only have one library for any given replicate then the merging step isnt carried out because the library-level and merged library-level BAM files will be exactly the same. +
+ Output files - Read duplicate marking is carried out using the Picard MarkDuplicates command. Duplicate reads are generally removed from the aligned reads to mitigate for fragments in the library that may have been sequenced more than once due to PCR biases. There is an option to keep duplicate reads with the `--keep_dups` parameter but its generally recommended to remove them to avoid the wrong interpretation of the results. A similar option has been provided to keep reads that are multi-mapped - `--keep_multi_map`. Other steps have been incorporated into the pipeline to filter the resulting alignments - see [`main README.md`](../README.md) for a more comprehensive listing, and the tools used at each step. +* `bwa/mergedLibrary/` + Merged library-level, coordinate sorted `*.bam` files after the marking of duplicates, and filtering based on various criteria. The file suffix for the final filtered files will be `*.mLb.clN.*`. If you specify the `--save_align_intermeds` parameter then two additional sets of files will be present. These represent the unfiltered alignments with duplicates marked (`*.mLb.mkD.*`), and in the case of paired-end datasets the filtered alignments before the removal of orphan read pairs (`*.mLb.flT.*`). +* `bwa/mergedLibrary/samtools_stats/` + SAMtools `*.flagstat`, `*.idxstats` and `*.stats` files generated from the alignment files. +* `bwa/mergedLibrary/picard_metrics/` + Alignment QC files from picard CollectMultipleMetrics and the metrics file from MarkDuplicates: `*_metrics` and `*.metrics.txt`, respectively. +* `bwa/mergedLibrary/picard_metrics/pdf/` + Alignment QC plot files in `*.pdf` format from picard CollectMultipleMetrics. +* `bwa/mergedLibrary/preseq/` + Preseq expected future yield file (`*.ccurve.txt`). - Certain cell types and tissues yield an enormous fraction (typically 20–80%) of unusable sequences of mitochondrial origin. This is a known problem that is specific to ATAC-seq library preps - see [Montefiori et al. 2017](https://www.nature.com/articles/s41598-017-02547-w). There is an option to keep these reads using the `--keep_mito` parameter but its generally recommended to remove these in order to get a more reliable assessment of the duplication rate from the rest of the genome, and to avoid any biases in the downstream analyses. +> **NB:** File names in the resulting directory (i.e. `bwa/mergedLibrary/`) will have the '`.mLb.`' suffix. - A selection of alignment-based QC metrics generated by Picard CollectMultipleMetrics and MarkDuplicates will be included in the MultiQC report. +
- ![MultiQC - Picard deduplication stats plot](images/mqc_picard_deduplication_plot.png) +![MultiQC - Picard deduplication stats plot](images/mqc_picard_deduplication_plot.png) - ![MultiQC - Picard insert size plot](images/mqc_picard_insert_size_plot.png) +![MultiQC - Picard insert size plot](images/mqc_picard_insert_size_plot.png) - The Preseq package is aimed at predicting and estimating the complexity of a genomic sequencing library, equivalent to predicting and estimating the number of redundant reads from a given sequencing depth and how many will be expected from additional sequencing using an initial sequencing experiment. The estimates can then be used to examine the utility of further sequencing, optimize the sequencing depth, or to screen multiple libraries to avoid low complexity samples. The dashed line shows a perfectly complex library where total reads = unique reads. +![MultiQC - Preseq library complexity plot](images/mqc_preseq_plot.png) - Note that these are predictive numbers only, not absolute. The MultiQC plot can sometimes give extreme sequencing depth on the X axis - click and drag from the left side of the plot to zoom in on more realistic numbers. +### Normalised bigWig files - ![MultiQC - Preseq library complexity plot](images/mqc_preseq_plot.png) +The [bigWig](https://genome.ucsc.edu/goldenpath/help/bigWig.html) format is in an indexed binary format useful for displaying dense, continuous data in Genome Browsers such as the [UCSC](https://genome.ucsc.edu/cgi-bin/hgTracks) and [IGV](http://software.broadinstitute.org/software/igv/). This mitigates the need to load the much larger BAM files for data visualisation purposes which will be slower and result in memory issues. The coverage values represented in the bigWig file can also be normalised in order to be able to compare the coverage across multiple samples - this is not possible with BAM files. The bigWig format is also supported by various bioinformatics software for downstream processing such as meta-profile plotting. - File names in the resulting directory (i.e. `bwa/mergedLibrary/`) will have the '`.mLb.`' suffix to denote **m**erged **L**i**b**raries. +
+ Output files - *Output directories*: - * `bwa/mergedLibrary/` - Merged library-level, coordinate sorted `*.bam` files after the marking of duplicates, and filtering based on various criteria. The file suffix for the final filtered files will be `*.mLb.clN.*`. If you specify the `--save_align_intermeds` parameter then two additional sets of files will be present. These represent the unfiltered alignments with duplicates marked (`*.mLb.mkD.*`), and in the case of paired-end datasets the filtered alignments before the removal of orphan read pairs (`*.mLb.flT.*`). - * `bwa/mergedLibrary/samtools_stats/` - SAMtools `*.flagstat`, `*.idxstats` and `*.stats` files generated from the alignment files. - * `bwa/mergedLibrary/picard_metrics/` - Alignment QC files from picard CollectMultipleMetrics and the metrics file from MarkDuplicates: `*_metrics` and `*.metrics.txt`, respectively. - * `bwa/mergedLibrary/picard_metrics/pdf/` - Alignment QC plot files in `*.pdf` format from picard CollectMultipleMetrics. - * `bwa/mergedLibrary/preseq/` - Preseq expected future yield file (`*.ccurve.txt`). +* `bwa/mergedLibrary/bigwig/` + Normalised `*.bigWig` files scaled to 1 million mapped reads. -2. **Normalised bigWig files** +
- *Documentation*: - [BEDTools](https://bedtools.readthedocs.io/en/latest/), [bedGraphToBigWig](https://genome.ucsc.edu/goldenpath/help/bigWig.html#Ex3) +### Coverage QC - *Description*: - The [bigWig](https://genome.ucsc.edu/goldenpath/help/bigWig.html) format is in an indexed binary format useful for displaying dense, continuous data in Genome Browsers such as the [UCSC](https://genome.ucsc.edu/cgi-bin/hgTracks) and [IGV](http://software.broadinstitute.org/software/igv/). This mitigates the need to load the much larger BAM files for data visualisation purposes which will be slower and result in memory issues. The coverage values represented in the bigWig file can also be normalised in order to be able to compare the coverage across multiple samples - this is not possible with BAM files. The bigWig format is also supported by various bioinformatics software for downstream processing such as meta-profile plotting. +[deepTools](https://deeptools.readthedocs.io/en/develop/content/list_of_tools.html) plotFingerprint is a useful QC for ATAC-seq data in order to see the relative enrichment of the samples in the experiment on a genome-wide basis (see [plotFingerprint docs](https://deeptools.readthedocs.io/en/develop/content/tools/plotFingerprint.html)). - *Output directories*: - * `bwa/mergedLibrary/bigwig/` - Normalised `*.bigWig` files scaled to 1 million mapped reads. +The results from deepTools plotProfile gives you a quick visualisation for the genome-wide enrichment of your samples at the TSS, and across the gene body. During the downstream analysis, you may want to refine the features/genes used to generate these plots in order to see a more specific condition-related effect. -3. **Coverage QC** +
+ Output files - *Documentation*: - [deepTools](https://deeptools.readthedocs.io/en/develop/content/list_of_tools.html) +* `bwa/mergedLibrary/deepTools/plotFingerprint/` + * Output files: `*.plotFingerprint.pdf`, `*.plotFingerprint.qcmetrics.txt`, `*.plotFingerprint.raw.txt` +* `bwa/mergedLibrary/deepTools/plotProfile/` + * Output files: `*.computeMatrix.mat.gz`, `*.computeMatrix.vals.mat.tab`, `*.plotProfile.pdf`, `*.plotProfile.tab`, `*.plotHeatmap.pdf`, `*.plotHeatmap.mat.tab`. - *Description*: - deepTools plotFingerprint is a useful QC for ATAC-seq data in order to see the relative enrichment of the samples in the experiment on a genome-wide basis (see [plotFingerprint docs](https://deeptools.readthedocs.io/en/develop/content/tools/plotFingerprint.html)). +
- ![MultiQC - deepTools plotFingerprint plot](images/mqc_deeptools_plotFingerprint_plot.png) +![MultiQC - deepTools plotFingerprint plot](images/mqc_deeptools_plotFingerprint_plot.png) - The results from deepTools plotProfile gives you a quick visualisation for the genome-wide enrichment of your samples at the TSS, and across the gene body. During the downstream analysis, you may want to refine the features/genes used to generate these plots in order to see a more specific condition-related effect. +![MultiQC - deepTools plotProfile plot](images/mqc_deeptools_plotProfile_plot.png) - ![MultiQC - deepTools plotProfile plot](images/mqc_deeptools_plotProfile_plot.png) +### Call peaks - *Output directories*: - * `bwa/mergedLibrary/deepTools/plotFingerprint/` - * Output files: `*.plotFingerprint.pdf`, `*.plotFingerprint.qcmetrics.txt`, `*.plotFingerprint.raw.txt` - * `bwa/mergedLibrary/deepTools/plotProfile/` - * Output files: `*.computeMatrix.mat.gz`, `*.computeMatrix.vals.mat.tab`, `*.plotProfile.pdf`, `*.plotProfile.tab`, `*.plotHeatmap.pdf`, `*.plotHeatmap.mat.tab`. +[MACS2](https://github.com/taoliu/MACS) is one of the most popular peak-calling algorithms for ChIP-seq data. For ATAC-seq data we are also looking for genome-wide regions of enrichment but in this case without comparison to a standard control sample (e.g. input DNA). By default, the peaks are called with the MACS2 `--broad` parameter. If, however, you would like to call narrow peaks then please provide the `--narrow_peak` parameter when running the pipeline. See [MACS2 outputs](https://github.com/taoliu/MACS#output-files) for a description of the output files generated by MACS2. -4. **Call peaks** +[HOMER annotatePeaks.pl](http://homer.ucsd.edu/homer/ngs/annotation.html) is used to annotate the peaks relative to known genomic features. HOMER is able to use the `--gtf` annotation file which is provided to the pipeline. Please note that some of the output columns will be blank because the annotation is not provided using HOMER's in-built database format. However, the more important fields required for downstream analysis will be populated i.e. *Annotation*, *Distance to TSS* and *Nearest Promoter ID*. - *Documentation*: - [MACS2](https://github.com/taoliu/MACS), [HOMER](http://homer.ucsd.edu/homer/ngs/annotation.html) +Various QC plots per sample including number of peaks, fold-change distribution, [FRiP score](https://genome.cshlp.org/content/22/9/1813.full.pdf+html) and peak-to-gene feature annotation are also generated by the pipeline. Where possible these have been integrated into the MultiQC report. - *Description*: - MACS2 is one of the most popular peak-calling algorithms for ChIP-seq data. For ATAC-seq data we are also looking for genome-wide regions of enrichment but in this case without comparison to a standard control sample (e.g. input DNA). By default, the peaks are called with the MACS2 `--broad` parameter. If, however, you would like to call narrow peaks then please provide the `--narrow_peak` parameter when running the pipeline. See [MACS2 outputs](https://github.com/taoliu/MACS#output-files) for a description of the output files generated by MACS2. +
+ Output files - ![MultiQC - MACS2 total peak count plot](images/mqc_macs2_peak_count_plot.png) +* `bwa/mergedLibrary/macs//` + * MACS2 output files: `*.xls`, `*.broadPeak` or `*.narrowPeak`, `*.gappedPeak` and `*summits.bed`. + The files generated will depend on whether MACS2 has been run in *narrowPeak* or *broadPeak* mode. + * HOMER peak-to-gene annotation file: `*.annotatePeaks.txt`. +* `bwa/mergedLibrary/macs//qc/` + * QC plots for MACS2 peaks: `macs_peak.plots.pdf` + * QC plots for peak-to-gene feature annotation: `macs_annotatePeaks.plots.pdf` + * MultiQC custom-content files for FRiP score, peak count and peak-to-gene ratios: `*.FRiP_mqc.tsv`, `*.count_mqc.tsv` and `macs_annotatePeaks.summary_mqc.tsv` respectively. - [HOMER annotatePeaks.pl](http://homer.ucsd.edu/homer/ngs/annotation.html) is used to annotate the peaks relative to known genomic features. HOMER is able to use the `--gtf` annotation file which is provided to the pipeline. Please note that some of the output columns will be blank because the annotation is not provided using HOMER's in-built database format. However, the more important fields required for downstream analysis will be populated i.e. *Annotation*, *Distance to TSS* and *Nearest Promoter ID*. +> **NB:** `` in the directory structure above corresponds to the type of peak that you have specified to call with MACS2 i.e. `broadPeak` or `narrowPeak`. If you so wish, you can call both narrow and broad peaks without redoing the preceding steps in the pipeline such as the alignment and filtering. For example, if you already have broad peaks then just add `--narrow_peak -resume` to the command you used to run the pipeline, and these will be called too! However, resuming the pipeline will only be possible if you have not deleted the `work/` directory generated by the pipeline. - ![MultiQC - HOMER annotatePeaks peak-to-gene feature ratio plot](images/mqc_annotatePeaks_feature_percentage_plot.png) +
- Various QC plots per sample including number of peaks, fold-change distribution, [FRiP score](https://genome.cshlp.org/content/22/9/1813.full.pdf+html) and peak-to-gene feature annotation are also generated by the pipeline. Where possible these have been integrated into the MultiQC report. +![MultiQC - MACS2 total peak count plot](images/mqc_macs2_peak_count_plot.png) - ![MultiQC - MACS2 peaks FRiP score plot](images/mqc_frip_score_plot.png) +![MultiQC - HOMER annotatePeaks peak-to-gene feature ratio plot](images/mqc_annotatePeaks_feature_percentage_plot.png) - `` in the directory structure below corresponds to the type of peak that you have specified to call with MACS2 i.e. `broadPeak` or `narrowPeak`. If you so wish, you can call both narrow and broad peaks without redoing the preceding steps in the pipeline such as the alignment and filtering. For example, if you already have broad peaks then just add `--narrow_peak -resume` to the command you used to run the pipeline, and these will be called too! However, resuming the pipeline will only be possible if you have not deleted the `work/` directory generated by the pipeline. +![MultiQC - MACS2 peaks FRiP score plot](images/mqc_frip_score_plot.png) - *Output directories*: - * `bwa/mergedLibrary/macs//` - * MACS2 output files: `*.xls`, `*.broadPeak` or `*.narrowPeak`, `*.gappedPeak` and `*summits.bed`. - The files generated will depend on whether MACS2 has been run in *narrowPeak* or *broadPeak* mode. - * HOMER peak-to-gene annotation file: `*.annotatePeaks.txt`. - * `bwa/mergedLibrary/macs//qc/` - * QC plots for MACS2 peaks: `macs_peak.plots.pdf` - * QC plots for peak-to-gene feature annotation: `macs_annotatePeaks.plots.pdf` - * MultiQC custom-content files for FRiP score, peak count and peak-to-gene ratios: `*.FRiP_mqc.tsv`, `*.count_mqc.tsv` and `macs_annotatePeaks.summary_mqc.tsv` respectively. +### Create and quantify consensus set of peaks -5. **Create and quantify consensus set of peaks** +In order to perform the differential binding analysis we need to be able to carry out the read quantification for the same intervals across **all** of the samples in the experiment. To this end, the individual peak-sets called per sample have to be merged together in order to create a consensus set of peaks. - *Documentation*: - [BEDTools](https://bedtools.readthedocs.io/en/latest/), [featureCounts](http://bioinf.wehi.edu.au/featureCounts/) +Using the consensus peaks it is possible to assess the degree of overlap between the peaks from a set of samples e.g. *Which consensus peaks contain peaks that are common/unique to a given set of samples?*. This may be useful for downstream filtering of peaks based on whether they are called in multiple replicates/conditions. Please note that it is possible for a consensus peak to contain multiple peaks from the same sample. Unfortunately, this is sample-dependent but the files generated by the pipeline do have columns that report such instances and allow you to factor them into any further analysis. - *Description*: - In order to perform the differential binding analysis we need to be able to carry out the read quantification for the same intervals across **all** of the samples in the experiment. To this end, the individual peak-sets called per sample have to be merged together in order to create a consensus set of peaks. +By default, the peak-sets are not filtered, therefore, the consensus peaks will be generated across the union set of peaks from all samples. However, you can increment the `--min_reps_consensus` parameter appropriately if you are confident you have good reproducibility amongst your replicates to create a "reproducible" set of consensus of peaks. In future iterations of the pipeline more formal analyses such as [IDR](https://projecteuclid.org/euclid.aoas/1318514284) may be implemented to obtain reproducible and high confidence peak-sets with which to perform this sort of analysis. - Using the consensus peaks it is possible to assess the degree of overlap between the peaks from a set of samples e.g. *Which consensus peaks contain peaks that are common/unique to a given set of samples?*. This may be useful for downstream filtering of peaks based on whether they are called in multiple replicates/conditions. Please note that it is possible for a consensus peak to contain multiple peaks from the same sample. Unfortunately, this is sample-dependent but the files generated by the pipeline do have columns that report such instances and allow you to factor them into any further analysis. +The [featureCounts](http://bioinf.wehi.edu.au/featureCounts/) tool is used to count the number of reads relative to the consensus peak-set across all of the samples. This essentially generates a file containing a matrix where the rows represent the consensus intervals, the columns represent all of the samples in the experiment, and the values represent the raw read counts. - By default, the peak-sets are not filtered, therefore, the consensus peaks will be generated across the union set of peaks from all samples. However, you can increment the `--min_reps_consensus` parameter appropriately if you are confident you have good reproducibility amongst your replicates to create a "reproducible" set of consensus of peaks. In future iterations of the pipeline more formal analyses such as [IDR](https://projecteuclid.org/euclid.aoas/1318514284) may be implemented to obtain reproducible and high confidence peak-sets with which to perform this sort of analysis. +
+ Output files - ![R - UpSetR peak intersection plot](images/r_upsetr_intersect_plot.png) +* `bwa/mergedLibrary/macs//consensus/` + * Consensus peak-set across all samples in `*.bed` format. + * Consensus peak-set across all samples in `*.saf` format. Required by featureCounts for read quantification. + * `.featureCounts.txt` file for read counts across all samples relative to consensus peak-set. + * HOMER `*.annotatePeaks.txt` peak-to-gene annotation file for consensus peaks. + * Spreadsheet representation of consensus peak-set across samples **with** gene annotation columns: `*.boolean.annotatePeaks.txt`. + The columns from individual peak files are included in this file along with the ability to filter peaks based on their presence or absence in multiple replicates/conditions. + * Spreadsheet representation of consensus peak-set across samples **without** gene annotation columns: `*.boolean.txt`. + Same as file above but without annotation columns. + * [UpSetR](https://cran.r-project.org/web/packages/UpSetR/README.html) files to illustrate peak intersection: `*.boolean.intersect.plot.pdf` and `*.boolean.intersect.txt`. - The featureCounts tool is used to count the number of reads relative to the consensus peak-set across all of the samples. This essentially generates a file containing a matrix where the rows represent the consensus intervals, the columns represent all of the samples in the experiment, and the values represent the raw read counts. +
- ![MultiQC - featureCounts consensus peak read assignment plot](images/mqc_featureCounts_assignment_plot.png) +![R - UpSetR peak intersection plot](images/r_upsetr_intersect_plot.png) - *Output directories*: - * `bwa/mergedLibrary/macs//consensus/` - * Consensus peak-set across all samples in `*.bed` format. - * Consensus peak-set across all samples in `*.saf` format. Required by featureCounts for read quantification. - * `.featureCounts.txt` file for read counts across all samples relative to consensus peak-set. - * HOMER `*.annotatePeaks.txt` peak-to-gene annotation file for consensus peaks. - * Spreadsheet representation of consensus peak-set across samples **with** gene annotation columns: `*.boolean.annotatePeaks.txt`. - The columns from individual peak files are included in this file along with the ability to filter peaks based on their presence or absence in multiple replicates/conditions. - * Spreadsheet representation of consensus peak-set across samples **without** gene annotation columns: `*.boolean.txt`. - Same as file above but without annotation columns. - * [UpSetR](https://cran.r-project.org/web/packages/UpSetR/README.html) files to illustrate peak intersection: `*.boolean.intersect.plot.pdf` and `*.boolean.intersect.txt`. +![MultiQC - featureCounts consensus peak read assignment plot](images/mqc_featureCounts_assignment_plot.png) -6. **Read counting and differential binding analysis** +### Read counting and differential binding analysis - *Documentation*: - [DESeq2](https://bioconductor.org/packages/release/bioc/vignettes/DESeq2/inst/doc/DESeq2.html), [R](https://www.r-project.org/) +[DESeq2](https://bioconductor.org/packages/release/bioc/vignettes/DESeq2/inst/doc/DESeq2.html) is more commonly used to perform differential expression analysis for RNA-seq datasets. However, it can also be used for ATAC-seq differential accessibility analysis, in which case you can imagine that instead of counts per gene for RNA-seq data we now have counts per accessible region. - *Description*: - DESeq2 is more commonly used to perform differential expression analysis for RNA-seq datasets. However, it can also be used for ATAC-seq differential accessibility analysis, in which case you can imagine that instead of counts per gene for RNA-seq data we now have counts per accessible region. +This pipeline uses a standardised DESeq2 analysis script to get an idea of the reproducibility within the experiment, and to assess the overall differential binding. Please note that this will not suit every experimental design, and if there are other problems with the experiment then it may not work as well as expected. For larger experiments, it may be recommended to use the `vst` transformation instead of the default `rlog` option. You can do this by providing the `--deseq2_vst` parameter to the pipeline. See [DESeq2 docs](http://bioconductor.org/packages/devel/bioc/vignettes/DESeq2/inst/doc/DESeq2.html#data-transformations-and-visualization) for a more detailed explanation. - This pipeline uses a standardised DESeq2 analysis script to get an idea of the reproducibility within the experiment, and to assess the overall differential binding. Please note that this will not suit every experimental design, and if there are other problems with the experiment then it may not work as well as expected. For larger experiments, it may be recommended to use the `vst` transformation instead of the default `rlog` option. You can do this by providing the `--deseq2_vst` parameter to the pipeline. See [DESeq2 docs](http://bioconductor.org/packages/devel/bioc/vignettes/DESeq2/inst/doc/DESeq2.html#data-transformations-and-visualization) for a more detailed explanation. +By default, all possible pairwise comparisons across the groups from a particular antibody (as defined in [`design.csv`](usage.md#--design)) are performed. The DESeq2 results are generated by the pipeline in various ways. You can load up the results across all of the comparisons in a single spreadsheet, or individual folders will also be created that contain the results specific to a particular comparison. For the latter, additional files will also be generated where the intervals have been pre-filtered based on a couple of standard FDR thresholds. Please see [DESeq2 output](http://bioconductor.org/packages/release/bioc/vignettes/DESeq2/inst/doc/DESeq2.html#differential-expression-analysis) for a description of the columns generated by DESeq2. - ![MultiQC - DESeq2 PCA plot](images/mqc_deseq2_pca_plot.png) +
+ Output files - ![MultiQC - DESeq2 sample similarity plot](images/mqc_deseq2_sample_similarity_plot.png) +* `bwa/mergedLibrary/macs//consensus//deseq2/` + * Differential binding `*.results.txt` spreadsheet containing results across all consensus peaks and all comparisons. + * `*.plots.pdf` file for PCA and hierarchical clustering. + * `*.log` file with information for number of differentially bound intervals at different FDR and fold-change thresholds for each comparison. + * `*.dds.rld.RData` file containing R `dds` and `rld` objects generated by DESeq2. + * `R_sessionInfo.log` file containing information about R, the OS and attached or loaded packages. +* `bwa/mergedLibrary/macs//consensus///` + * `*.results.txt` spreadsheet containing comparison-specific DESeq2 output for differential binding results across all peaks. + * Subset of above file for peaks that pass FDR <= 0.01 (`*FDR0.01.results.txt`) and FDR <= 0.05 (`*FDR0.05.results.txt`). + * BED files for peaks that pass FDR <= 0.01 (`*FDR0.01.results.bed`) and FDR <= 0.05 (`*FDR0.05.results.bed`). + * MA, Volcano, clustering and scatterplots at FDR <= 0.01 and FDR <= 0.05: `*deseq2.plots.pdf`. +* `bwa/mergedLibrary/macs//consensus//sizeFactors/` + Files containing DESeq2 sizeFactors per sample: `*.txt` and `*.RData`. - By default, all possible pairwise comparisons across the groups from a particular antibody (as defined in [`design.csv`](usage.md#--design)) are performed. The DESeq2 results are generated by the pipeline in various ways. You can load up the results across all of the comparisons in a single spreadsheet, or individual folders will also be created that contain the results specific to a particular comparison. For the latter, additional files will also be generated where the intervals have been pre-filtered based on a couple of standard FDR thresholds. Please see [DESeq2 output](http://bioconductor.org/packages/release/bioc/vignettes/DESeq2/inst/doc/DESeq2.html#differential-expression-analysis) for a description of the columns generated by DESeq2. +
- ![R - DESeq2 MA plot](images/r_deseq2_ma_plot.png) +![MultiQC - DESeq2 PCA plot](images/mqc_deseq2_pca_plot.png) - ![R - DESeq2 Volcano plot](images/r_deseq2_volcano_plot.png) +![MultiQC - DESeq2 sample similarity plot](images/mqc_deseq2_sample_similarity_plot.png) - *Output directories*: - * `bwa/mergedLibrary/macs//consensus//deseq2/` - * Differential binding `*.results.txt` spreadsheet containing results across all consensus peaks and all comparisons. - * `*.plots.pdf` file for PCA and hierarchical clustering. - * `*.log` file with information for number of differentially bound intervals at different FDR and fold-change thresholds for each comparison. - * `*.dds.rld.RData` file containing R `dds` and `rld` objects generated by DESeq2. - * `R_sessionInfo.log` file containing information about R, the OS and attached or loaded packages. - * `bwa/mergedLibrary/macs//consensus///` - * `*.results.txt` spreadsheet containing comparison-specific DESeq2 output for differential binding results across all peaks. - * Subset of above file for peaks that pass FDR <= 0.01 (`*FDR0.01.results.txt`) and FDR <= 0.05 (`*FDR0.05.results.txt`). - * BED files for peaks that pass FDR <= 0.01 (`*FDR0.01.results.bed`) and FDR <= 0.05 (`*FDR0.05.results.bed`). - * MA, Volcano, clustering and scatterplots at FDR <= 0.01 and FDR <= 0.05: `*deseq2.plots.pdf`. - * `bwa/mergedLibrary/macs//consensus//sizeFactors/` - Files containing DESeq2 sizeFactors per sample: `*.txt` and `*.RData`. +![R - DESeq2 MA plot](images/r_deseq2_ma_plot.png) -7. **ataqv** +![R - DESeq2 Volcano plot](images/r_deseq2_volcano_plot.png) - *Software*: - [ataqv](https://parkerlab.github.io/ataqv/) +### ataqv - *Description*: - ataqv is a toolkit for measuring and comparing ATAC-seq results. It was written to help understand how well ATAC-seq assays have worked, and to make it easier to spot differences that might be caused by library prep or sequencing. +[ataqv](https://parkerlab.github.io/ataqv/) is a toolkit for measuring and comparing ATAC-seq results. It was written to help understand how well ATAC-seq assays have worked, and to make it easier to spot differences that might be caused by library prep or sequencing. Please see [ataqv homepage](https://parkerlab.github.io/ataqv/) for documentation and an example report. - Please see [ataqv homepage](https://parkerlab.github.io/ataqv/) for documentation and an example report. +
+ Output files - *Output directories*: - * `bwa/mergedLibrary/ataqv//` - `.json` files containing ATAC-seq specific metrics for each sample. - * `bwa/mergedLibrary/ataqv//html/` - Folder containing ataqv results aggregated across all samples for visualisation via an internet browser. +* `bwa/mergedLibrary/ataqv//` + `.json` files containing ATAC-seq specific metrics for each sample. +* `bwa/mergedLibrary/ataqv//html/` + Folder containing ataqv results aggregated across all samples for visualisation via an internet browser. + +
## Merged replicate-level analysis The alignments associated with all of the replicates from the same experimental condition can also be merged. This can be useful to increase the coverage for peak-calling and for other analyses that require high sequencing depth such as [motif footprinting](https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3959825/). The analysis steps and directory structure for `bwa/mergedLibrary/` and `bwa/mergedReplicate/` are almost identical. -File names in the resulting directory (i.e. `bwa/mergedReplicate/`) will have the '`.mRp.`' suffix to denote **m**erged **R**e**p**licates. +File names in the resulting directory (i.e. `bwa/mergedReplicate/`) will have the '`.mRp.`' suffix. You can skip this portion of the analysis by specifying the `--skip_merge_replicates` parameter. ->NB: Merged library-level alignments will be used for read counting relative to the consensus merged replicate-level peakset. This is the only way in which differential analysis can be performed at the merged replicate-level. +> **NB:** Merged library-level alignments will be used for read counting relative to the consensus merged replicate-level peakset. This is the only way in which differential analysis can be performed at the merged replicate-level. ## Aggregate analysis -1. **Present QC for the raw read, alignment, peak and differential accessibility results** +### Present QC for the raw read, alignment, peak and differential accessibility results + +[MultiQC](https://multiqc.info/docs/) is a visualisation tool that generates a single HTML report summarising all samples in your project. Most of the pipeline QC results are visualised in the report and further statistics are available within the report data directory. + +Results generated by MultiQC collate pipeline QC from FastQC, TrimGalore, samtools flagstat, samtools idxstats, samtools stats, picard CollectMultipleMetrics, picard MarkDuplicates, Preseq, deepTools plotProfile, deepTools plotFingerprint and featureCounts. The default [`multiqc config file`](../assets/multiqc_config.yaml) also contains the provision for loading custom-content to report peak counts, FRiP scores, peak-to-gene annnotation proportions, sample-similarity heatmaps and PCA plots. - *Documentation*: - [MultiQC](https://multiqc.info/docs/) +The pipeline has special steps which also allow the software versions to be reported in the MultiQC output for future traceability. For more information about how to use MultiQC reports, see . - *Description*: - MultiQC is a visualisation tool that generates a single HTML report summarising all samples in your project. Most of the pipeline QC results are visualised in the report and further statistics are available within the report data directory. +
+ Output files - Results generated by MultiQC collate pipeline QC from FastQC, TrimGalore, samtools flagstat, samtools idxstats, samtools stats, picard CollectMultipleMetrics, picard MarkDuplicates, Preseq, deepTools plotProfile, deepTools plotFingerprint and featureCounts. The default [`multiqc config file`](../assets/multiqc_config.yaml) also contains the provision for loading custom-content to report peak counts, FRiP scores, peak-to-gene annnotation proportions, sample-similarity heatmaps and PCA plots. +* `multiqc//` + * `multiqc_report.html` - a standalone HTML file that can be viewed in your web browser. + * `multiqc_data/` - directory containing parsed statistics from the different tools used in the pipeline. + * `multiqc_plots/` - directory containing static images from the report in various formats. - The pipeline has special steps which also allow the software versions to be reported in the MultiQC output for future traceability. For more information about how to use MultiQC reports, see . +
- *Output directories*: - * `multiqc//` - * `multiqc_report.html` - a standalone HTML file that can be viewed in your web browser. - * `multiqc_data/` - directory containing parsed statistics from the different tools used in the pipeline. - * `multiqc_plots/` - directory containing static images from the report in various formats. +### Create IGV session file -2. **Create IGV session file** +An [IGV](https://software.broadinstitute.org/software/igv/UserGuide) session file will be created at the end of the pipeline containing the normalised bigWig tracks, per-sample peaks, consensus peaks and differential sites. This avoids having to load all of the data individually into IGV for visualisation. - *Documentation*: - [IGV](https://software.broadinstitute.org/software/igv/UserGuide) +The genome fasta file required for the IGV session will be the same as the one that was provided to the pipeline. This will be copied into `genome/` to overcome any loading issues. If you prefer to use another path or an in-built genome provided by IGV just change the `genome` entry in the second-line of the session file. - *Description*: - An IGV session file will be created at the end of the pipeline containing the normalised bigWig tracks, per-sample peaks, consensus peaks and differential sites. This avoids having to load all of the data individually into IGV for visualisation. +The file paths in the IGV session file will only work if the results are kept in the same place on your storage. If the results are moved or for example, if you prefer to load the data over the web then just replace the file paths with others that are more appropriate. - The genome fasta file required for the IGV session will be the same as the one that was provided to the pipeline. This will be copied into `genome/` to overcome any loading issues. If you prefer to use another path or an in-built genome provided by IGV just change the `genome` entry in the second-line of the session file. +Once installed, open IGV, go to `File > Open Session` and select the `igv_session.xml` file for loading. - The file paths in the IGV session file will only work if the results are kept in the same place on your storage. If the results are moved or for example, if you prefer to load the data over the web then just replace the file paths with others that are more appropriate. +> **NB:** If you are not using an in-built genome provided by IGV you will need to load the annotation yourself e.g. in .gtf and/or .bed format. - Once installed, open IGV, go to `File > Open Session` and select the `igv_session.xml` file for loading. +
+ Output files - >NB: If you are not using an in-built genome provided by IGV you will need to load the annotation yourself e.g. in .gtf and/or .bed format. +* `igv//` + * `igv_session.xml` file. + * `igv_files.txt` file containing a listing of the files used to create the IGV session, and their allocated colours. - ![IGV screenshot](images/igv_screenshot.png) +
- *Output directories*: - * `igv//` - * `igv_session.xml` file. - * `igv_files.txt` file containing a listing of the files used to create the IGV session, and their allocated colours. +![IGV screenshot](images/igv_screenshot.png) ## Other results -1. **Reference genome files** +### Reference genome files + +Reference genome-specific files can be useful to keep for the downstream processing of the results. + +
+ Output files - *Documentation*: - [BWA](https://sourceforge.net/projects/bio-bwa/files/), [BEDTools](https://bedtools.readthedocs.io/en/latest/), [SAMtools](http://samtools.sourceforge.net/) +* `genome/` + A number of genome-specific files are generated by the pipeline in order to aid in the filtering of the data, and because they are required by standard tools such as BEDTools. These can be found in this directory along with the genome fasta file which is required by IGV. If using a genome from AWS iGenomes and if it exists a `README.txt` file containing information about the annotation version will also be saved in this directory. +* `genome/BWAIndex/` + If the `--save_reference` parameter is provided then the alignment indices generated by the pipeline will be saved in this directory. This can be quite a time-consuming process so it permits their reuse for future runs of the pipeline or for other purposes. - *Description*: - Reference genome-specific files can be useful to keep for the downstream processing of the results. +
- *Output directories*: - * `genome/` - A number of genome-specific files are generated by the pipeline in order to aid in the filtering of the data, and because they are required by standard tools such as BEDTools. These can be found in this directory along with the genome fasta file which is required by IGV. If using a genome from AWS iGenomes and if it exists a `README.txt` file containing information about the annotation version will also be saved in this directory. - * `genome/BWAIndex/` - If the `--save_reference` parameter is provided then the alignment indices generated by the pipeline will be saved in this directory. This can be quite a time-consuming process so it permits their reuse for future runs of the pipeline or for other purposes. +### Pipeline information -2. **Pipeline information** +[Nextflow](https://www.nextflow.io/docs/latest/tracing.html) provides excellent functionality for generating various reports relevant to the running and execution of the pipeline. This will allow you to trouble-shoot errors with the running of the pipeline, and also provide you with other information such as launch commands, run times and resource usage. - *Documentation*: - [Nextflow!](https://www.nextflow.io/docs/latest/tracing.html) +
+ Output files - *Description*: - Nextflow provides excellent functionality for generating various reports relevant to the running and execution of the pipeline. This will allow you to trouble-shoot errors with the running of the pipeline, and also provide you with other information such as launch commands, run times and resource usage. +* `pipeline_info/` + * Reports generated by the pipeline - `pipeline_report.html`, `pipeline_report.txt` and `software_versions.csv`. + * Reports generated by Nextflow - `execution_report.html`, `execution_timeline.html`, `execution_trace.txt` and `pipeline_dag.svg`. + * Reformatted design files used as input to the pipeline - `design_reads.csv` and `design_controls.csv`. +* `Documentation/` + Documentation for interpretation of results in HTML format - `results_description.html`. - *Output directories*: - * `pipeline_info/` - * Reports generated by the pipeline - `pipeline_report.html`, `pipeline_report.txt` and `software_versions.csv`. - * Reports generated by Nextflow - `execution_report.html`, `execution_timeline.html`, `execution_trace.txt` and `pipeline_dag.svg`. - * Reformatted design files used as input to the pipeline - `design_reads.csv` and `design_controls.csv`. - * `Documentation/` - Documentation for interpretation of results in HTML format - `results_description.html`. +
From 65fba3a1746047be8cc8bbccb8fd1eaf5820d78a Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 30 Jun 2020 14:51:58 +0100 Subject: [PATCH 205/226] Fix markdownlint --- CHANGELOG.md | 26 ++++----- docs/usage.md | 148 +++++++++++++++++++++++++------------------------- 2 files changed, 87 insertions(+), 87 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 1a13e46e..7a79a3c8 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -19,11 +19,11 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * Replace `set` with `tuple` and `file()` with `path()` in all processes * Capitalise process names * Parameters: - * `--skip_peak_qc` to skip MACS2 peak QC plot generation - * `--skip_peak_annotation` to skip annotation of MACS2 and consensus peaks with HOMER - * `--skip_consensus_peaks` to skip consensus peak generation - * `--deseq2_vst` to use variance stabilizing transformation (VST) instead of regularized log transformation (rlog) with DESeq2 - * `--publish_dir_mode` to customise method of publishing results to output directory [nf-core/tools#585](https://github.com/nf-core/tools/issues/585) + * `--skip_peak_qc` to skip MACS2 peak QC plot generation + * `--skip_peak_annotation` to skip annotation of MACS2 and consensus peaks with HOMER + * `--skip_consensus_peaks` to skip consensus peak generation + * `--deseq2_vst` to use variance stabilizing transformation (VST) instead of regularized log transformation (rlog) with DESeq2 + * `--publish_dir_mode` to customise method of publishing results to output directory [nf-core/tools#585](https://github.com/nf-core/tools/issues/585) ### `Fixed` @@ -83,14 +83,14 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * Add `CITATIONS.md` file * Capitalised process names * Add parameters: - * `--seq_center` - * `--trim_nextseq` - * `--fingerprint_bins` - * `--broad_cutoff` - * `--min_reps_consensus` - * `--save_macs_pileup` - * `--skip_diff_analysis` - * `--skip_*` for skipping QC steps + * `--seq_center` + * `--trim_nextseq` + * `--fingerprint_bins` + * `--broad_cutoff` + * `--min_reps_consensus` + * `--save_macs_pileup` + * `--skip_diff_analysis` + * `--skip_*` for skipping QC steps ### `Fixed` diff --git a/docs/usage.md b/docs/usage.md index b8252113..41eff287 100755 --- a/docs/usage.md +++ b/docs/usage.md @@ -5,73 +5,73 @@ * [Table of contents](#table-of-contents) * [Introduction](#introduction) * [Running the pipeline](#running-the-pipeline) - * [Updating the pipeline](#updating-the-pipeline) - * [Reproducibility](#reproducibility) + * [Updating the pipeline](#updating-the-pipeline) + * [Reproducibility](#reproducibility) * [Main arguments](#main-arguments) - * [`-profile`](#-profile) - * [`--input`](#--input) + * [`-profile`](#-profile) + * [`--input`](#--input) * [Generic arguments](#generic-arguments) - * [`--single_end`](#--single_end) - * [`--seq_center`](#--seq_center) - * [`--fragment_size`](#--fragment_size) - * [`--fingerprint_bins`](#--fingerprint_bins) + * [`--single_end`](#--single_end) + * [`--seq_center`](#--seq_center) + * [`--fragment_size`](#--fragment_size) + * [`--fingerprint_bins`](#--fingerprint_bins) * [Reference genomes](#reference-genomes) - * [`--genome` (using iGenomes)](#--genome-using-igenomes) - * [`--fasta`](#--fasta) - * [`--gtf`](#--gtf) - * [`--bwa_index`](#--bwa_index) - * [`--gene_bed`](#--gene_bed) - * [`--tss_bed`](#--tss_bed) - * [`--macs_gsize`](#--macs_gsize) - * [`--blacklist`](#--blacklist) - * [`--mito_name`](#--mito_name) - * [`--save_reference`](#--save_reference) - * [`--igenomes_ignore`](#--igenomes_ignore) + * [`--genome` (using iGenomes)](#--genome-using-igenomes) + * [`--fasta`](#--fasta) + * [`--gtf`](#--gtf) + * [`--bwa_index`](#--bwa_index) + * [`--gene_bed`](#--gene_bed) + * [`--tss_bed`](#--tss_bed) + * [`--macs_gsize`](#--macs_gsize) + * [`--blacklist`](#--blacklist) + * [`--mito_name`](#--mito_name) + * [`--save_reference`](#--save_reference) + * [`--igenomes_ignore`](#--igenomes_ignore) * [Adapter trimming](#adapter-trimming) - * [`--skip_trimming`](#--skip_trimming) - * [`--save_trimmed`](#--save_trimmed) + * [`--skip_trimming`](#--skip_trimming) + * [`--save_trimmed`](#--save_trimmed) * [Alignments](#alignments) - * [`--keep_mito`](#--keep_mito) - * [`--keep_dups`](#--keep_dups) - * [`--keep_multi_map`](#--keep_multi_map) - * [`--skip_merge_replicates`](#--skip_merge_replicates) - * [`--save_align_intermeds`](#--save_align_intermeds) + * [`--keep_mito`](#--keep_mito) + * [`--keep_dups`](#--keep_dups) + * [`--keep_multi_map`](#--keep_multi_map) + * [`--skip_merge_replicates`](#--skip_merge_replicates) + * [`--save_align_intermeds`](#--save_align_intermeds) * [Peaks](#peaks) - * [`--narrow_peak`](#--narrow_peak) - * [`--broad_cutoff`](#--broad_cutoff) - * [`--min_reps_consensus`](#--min_reps_consensus) - * [`--save_macs_pileup`](#--save_macs_pileup) - * [`--skip_peak_qc`](#--skip_peak_qc) - * [`--skip_peak_annotation`](#--skip_peak_annotation) - * [`--skip_consensus_peaks`](#--skip_consensus_peaks) + * [`--narrow_peak`](#--narrow_peak) + * [`--broad_cutoff`](#--broad_cutoff) + * [`--min_reps_consensus`](#--min_reps_consensus) + * [`--save_macs_pileup`](#--save_macs_pileup) + * [`--skip_peak_qc`](#--skip_peak_qc) + * [`--skip_peak_annotation`](#--skip_peak_annotation) + * [`--skip_consensus_peaks`](#--skip_consensus_peaks) * [Differential analysis](#differential_analysis) - * [`--deseq2_vst`](#--deseq2_vst) - * [`--skip_diff_analysis`](#--skip_diff_analysis) + * [`--deseq2_vst`](#--deseq2_vst) + * [`--skip_diff_analysis`](#--skip_diff_analysis) * [Skipping QC steps](#skipping-qc-steps) * [Job resources](#job-resources) - * [Automatic resubmission](#automatic-resubmission) - * [Custom resource requests](#custom-resource-requests) + * [Automatic resubmission](#automatic-resubmission) + * [Custom resource requests](#custom-resource-requests) * [AWS Batch specific parameters](#aws-batch-specific-parameters) - * [`--awsqueue`](#--awsqueue) - * [`--awsregion`](#--awsregion) - * [`--awscli`](#--awscli) + * [`--awsqueue`](#--awsqueue) + * [`--awsregion`](#--awsregion) + * [`--awscli`](#--awscli) * [Other command line parameters](#other-command-line-parameters) - * [`--outdir`](#--outdir) - * [`--publish_dir_mode`](#--publish_dir_mode) - * [`--email`](#--email) - * [`--email_on_fail`](#--email_on_fail) - * [`--max_multiqc_email_size`](#--max_multiqc_email_size) - * [`-name`](#-name) - * [`-resume`](#-resume) - * [`-c`](#-c) - * [`--custom_config_version`](#--custom_config_version) - * [`--custom_config_base`](#--custom_config_base) - * [`--max_memory`](#--max_memory) - * [`--max_time`](#--max_time) - * [`--max_cpus`](#--max_cpus) - * [`--plaintext_email`](#--plaintext_email) - * [`--monochrome_logs`](#--monochrome_logs) - * [`--multiqc_config`](#--multiqc_config) + * [`--outdir`](#--outdir) + * [`--publish_dir_mode`](#--publish_dir_mode) + * [`--email`](#--email) + * [`--email_on_fail`](#--email_on_fail) + * [`--max_multiqc_email_size`](#--max_multiqc_email_size) + * [`-name`](#-name) + * [`-resume`](#-resume) + * [`-c`](#-c) + * [`--custom_config_version`](#--custom_config_version) + * [`--custom_config_base`](#--custom_config_base) + * [`--max_memory`](#--max_memory) + * [`--max_time`](#--max_time) + * [`--max_cpus`](#--max_cpus) + * [`--plaintext_email`](#--plaintext_email) + * [`--monochrome_logs`](#--monochrome_logs) + * [`--multiqc_config`](#--multiqc_config) ## Introduction @@ -136,18 +136,18 @@ They are loaded in sequence, so later profiles can overwrite earlier profiles. If `-profile` is not specified, the pipeline will run locally and expect all software to be installed and available on the `PATH`. This is _not_ recommended. * `docker` - * A generic configuration profile to be used with [Docker](http://docker.com/) - * Pulls software from dockerhub: [`nfcore/atacseq`](http://hub.docker.com/r/nfcore/atacseq/) + * A generic configuration profile to be used with [Docker](http://docker.com/) + * Pulls software from dockerhub: [`nfcore/atacseq`](http://hub.docker.com/r/nfcore/atacseq/) * `singularity` - * A generic configuration profile to be used with [Singularity](http://singularity.lbl.gov/) - * Pulls software from DockerHub: [`nfcore/atacseq`](http://hub.docker.com/r/nfcore/atacseq/) + * A generic configuration profile to be used with [Singularity](http://singularity.lbl.gov/) + * Pulls software from DockerHub: [`nfcore/atacseq`](http://hub.docker.com/r/nfcore/atacseq/) * `conda` - * Please only use Conda as a last resort i.e. when it's not possible to run the pipeline with Docker or Singularity. - * A generic configuration profile to be used with [Conda](https://conda.io/docs/) - * Pulls most software from [Bioconda](https://bioconda.github.io/) + * Please only use Conda as a last resort i.e. when it's not possible to run the pipeline with Docker or Singularity. + * A generic configuration profile to be used with [Conda](https://conda.io/docs/) + * Pulls most software from [Bioconda](https://bioconda.github.io/) * `test` - * A profile with a complete configuration for automated testing - * Includes links to test data so needs no other parameters + * A profile with a complete configuration for automated testing + * Includes links to test data so needs no other parameters ### `--input` @@ -235,13 +235,13 @@ There are 31 different species supported in the iGenomes references. To run the You can find the keys to specify the genomes in the [iGenomes config file](../conf/igenomes.config). Common genomes that are supported are: * Human - * `--genome GRCh37` + * `--genome GRCh37` * Mouse - * `--genome GRCm38` + * `--genome GRCm38` * _Drosophila_ - * `--genome BDGP6` + * `--genome BDGP6` * _S. cerevisiae_ - * `--genome 'R64-1-1'` + * `--genome 'R64-1-1'` > There are numerous others - check the config file for more. @@ -338,15 +338,15 @@ The pipeline accepts a number of parameters to change how the trimming is done, You can specify custom trimming parameters as follows: * `--clip_r1 [int]` - * Instructs Trim Galore to remove [int] bp from the 5' end of read 1 (for single-end reads). + * Instructs Trim Galore to remove [int] bp from the 5' end of read 1 (for single-end reads). * `--clip_r2 [int]` - * Instructs Trim Galore to remove [int] bp from the 5' end of read 2 (paired-end reads only). + * Instructs Trim Galore to remove [int] bp from the 5' end of read 2 (paired-end reads only). * `--three_prime_clip_r1 [int]` - * Instructs Trim Galore to remove [int] bp from the 3' end of read 1 _AFTER_ adapter/quality trimming has been + * Instructs Trim Galore to remove [int] bp from the 3' end of read 1 _AFTER_ adapter/quality trimming has been * `--three_prime_clip_r2 [int]` - * Instructs Trim Galore to remove [int] bp from the 3' end of read 2 _AFTER_ adapter/quality trimming has been performed. + * Instructs Trim Galore to remove [int] bp from the 3' end of read 2 _AFTER_ adapter/quality trimming has been performed. * `--trim_nextseq [int]` - * This enables the option Cutadapt `--nextseq-trim=3'CUTOFF` option via Trim Galore, which will set a quality cutoff (that is normally given with -q instead), but qualities of G bases are ignored. This trimming is in common for the NextSeq- and NovaSeq-platforms, where basecalls without any signal are called as high-quality G bases. + * This enables the option Cutadapt `--nextseq-trim=3'CUTOFF` option via Trim Galore, which will set a quality cutoff (that is normally given with -q instead), but qualities of G bases are ignored. This trimming is in common for the NextSeq- and NovaSeq-platforms, where basecalls without any signal are called as high-quality G bases. ### `--skip_trimming` From 52909121ea38d42a1ba9d0a59879d20949fe03fa Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 30 Jun 2020 16:14:32 +0100 Subject: [PATCH 206/226] Remove trailing spaces --- docs/output.md | 195 ++++++++++++++++++++++++------------------------- 1 file changed, 96 insertions(+), 99 deletions(-) diff --git a/docs/output.md b/docs/output.md index db87dab3..058ab454 100755 --- a/docs/output.md +++ b/docs/output.md @@ -16,15 +16,15 @@ The initial QC and alignments are performed at the library-level e.g. if the sam ### Raw read QC -[FastQC](http://www.bioinformatics.babraham.ac.uk/projects/fastqc/Help/) gives general quality metrics about your reads. It provides information about the quality score distribution across your reads, the per base sequence content (%A/C/G/T). You get information about adapter contamination and other overrepresented sequences. +[FastQC](http://www.bioinformatics.babraham.ac.uk/projects/fastqc/Help/) gives general quality metrics about your reads. It provides information about the quality score distribution across your reads, the per base sequence content (%A/C/G/T). You get information about adapter contamination and other overrepresented sequences.
Output files -* `fastqc/` -FastQC `*.html` files for read 1 (*and read2 if paired-end*) **before** adapter trimming. -* `fastqc/zips/` -FastQC `*.zip` files for read 1 (*and read2 if paired-end*) **before** adapter trimming. +* `fastqc/` + * `*_fastqc.html`: FastQC report containing quality metrics for read 1 (*and read2 if paired-end*) **before** adapter trimming. +* `fastqc/zips/` + * `*_fastqc.zip`: Zip archive containing the FastQC report, tab-delimited data file and plot images.
@@ -35,14 +35,14 @@ FastQC `*.zip` files for read 1 (*and read2 if paired-end*) **before** adapter t
Output files -* `trim_galore/` - If `--save_trimmed` is specified FastQ files **after** adapter trimming will be placed in this directory. -* `trim_galore/logs/` - `*.log` files generated by Trim Galore!. -* `trim_galore/fastqc/` - FastQC `*.html` files for read 1 (*and read2 if paired-end*) **after** adapter trimming. -* `trim_galore/fastqc/zips/` - FastQC `*.zip` files for read 1 (*and read2 if paired-end*) **after** adapter trimming. +* `trim_galore/` + * `*fastq.gz`: If `--save_trimmed` is specified, FastQ files **after** adapter trimming will be placed in this directory. +* `trim_galore/logs/` + * `*.log`: Log file generated by Trim Galore!. +* `trim_galore/fastqc/` + * `*_fastqc.html`: FastQC report containing quality metrics for read 1 (*and read2 if paired-end*) **after** adapter trimming. +* `trim_galore/fastqc/zips/` + * `*_fastqc.zip`: Zip archive containing the FastQC report, tab-delimited data file and plot images.
@@ -56,9 +56,9 @@ Adapter-trimmed reads are mapped to the reference assembly using [BWA](http://bi Output files * `bwa/library/` - The files resulting from the alignment of individual libraries are not saved by default so this directory will not be present in your results. You can override this behaviour with the use of the `--save_align_intermeds` flag in which case it will contain the coordinate sorted alignment files in [`*.bam`](https://samtools.github.io/hts-specs/SAMv1.pdf) format. -* `bwa/library/samtools_stats/` - SAMtools `*.flagstat`, `*.idxstats` and `*.stats` files generated from the alignment files. + * `*.bam`: The files resulting from the alignment of individual libraries are not saved by default so this directory will not be present in your results. You can override this behaviour with the use of the `--save_align_intermeds` flag in which case it will contain the coordinate sorted alignment files in [`*.bam`](https://samtools.github.io/hts-specs/SAMv1.pdf) format. +* `bwa/library/samtools_stats/` + * SAMtools `*.flagstat`, `*.idxstats` and `*.stats` files generated from the alignment files. > **NB:** File names in the resulting directory (i.e. `bwa/library/`) will have the '`.Lb.`' suffix. @@ -85,16 +85,17 @@ The [Preseq](http://smithlabresearch.org/software/preseq/) package is aimed at p
Output files -* `bwa/mergedLibrary/` - Merged library-level, coordinate sorted `*.bam` files after the marking of duplicates, and filtering based on various criteria. The file suffix for the final filtered files will be `*.mLb.clN.*`. If you specify the `--save_align_intermeds` parameter then two additional sets of files will be present. These represent the unfiltered alignments with duplicates marked (`*.mLb.mkD.*`), and in the case of paired-end datasets the filtered alignments before the removal of orphan read pairs (`*.mLb.flT.*`). -* `bwa/mergedLibrary/samtools_stats/` - SAMtools `*.flagstat`, `*.idxstats` and `*.stats` files generated from the alignment files. -* `bwa/mergedLibrary/picard_metrics/` - Alignment QC files from picard CollectMultipleMetrics and the metrics file from MarkDuplicates: `*_metrics` and `*.metrics.txt`, respectively. -* `bwa/mergedLibrary/picard_metrics/pdf/` - Alignment QC plot files in `*.pdf` format from picard CollectMultipleMetrics. -* `bwa/mergedLibrary/preseq/` - Preseq expected future yield file (`*.ccurve.txt`). +* `bwa/mergedLibrary/` + * `*.bam`: Merged library-level, coordinate sorted `*.bam` files after the marking of duplicates, and filtering based on various criteria. The file suffix for the final filtered files will be `*.mLb.clN.*`. If you specify the `--save_align_intermeds` parameter then two additional sets of files will be present. These represent the unfiltered alignments with duplicates marked (`*.mLb.mkD.*`), and in the case of paired-end datasets the filtered alignments before the removal of orphan read pairs (`*.mLb.flT.*`). +* `bwa/mergedLibrary/samtools_stats/` + * SAMtools `*.flagstat`, `*.idxstats` and `*.stats` files generated from the alignment files. +* `bwa/mergedLibrary/picard_metrics/` + * `*_metrics`: Alignment QC files from picard CollectMultipleMetrics. + * `*.metrics.txt`: Metrics file from MarkDuplicates. +* `bwa/mergedLibrary/picard_metrics/pdf/` + * `*.pdf`: Alignment QC plot files from picard CollectMultipleMetrics. +* `bwa/mergedLibrary/preseq/` + * `*.ccurve.txt`: Preseq expected future yield file. > **NB:** File names in the resulting directory (i.e. `bwa/mergedLibrary/`) will have the '`.mLb.`' suffix. @@ -102,9 +103,9 @@ The [Preseq](http://smithlabresearch.org/software/preseq/) package is aimed at p ![MultiQC - Picard deduplication stats plot](images/mqc_picard_deduplication_plot.png) -![MultiQC - Picard insert size plot](images/mqc_picard_insert_size_plot.png) +![MultiQC - Picard insert size plot](images/mqc_picard_insert_size_plot.png) -![MultiQC - Preseq library complexity plot](images/mqc_preseq_plot.png) +![MultiQC - Preseq library complexity plot](images/mqc_preseq_plot.png) ### Normalised bigWig files @@ -113,8 +114,8 @@ The [bigWig](https://genome.ucsc.edu/goldenpath/help/bigWig.html) format is in a
Output files -* `bwa/mergedLibrary/bigwig/` - Normalised `*.bigWig` files scaled to 1 million mapped reads. +* `bwa/mergedLibrary/bigwig/` + * `*.bigWig`: Normalised bigWig files scaled to 1 million mapped reads.
@@ -127,10 +128,10 @@ The results from deepTools plotProfile gives you a quick visualisation for the g
Output files -* `bwa/mergedLibrary/deepTools/plotFingerprint/` - * Output files: `*.plotFingerprint.pdf`, `*.plotFingerprint.qcmetrics.txt`, `*.plotFingerprint.raw.txt` -* `bwa/mergedLibrary/deepTools/plotProfile/` - * Output files: `*.computeMatrix.mat.gz`, `*.computeMatrix.vals.mat.tab`, `*.plotProfile.pdf`, `*.plotProfile.tab`, `*.plotHeatmap.pdf`, `*.plotHeatmap.mat.tab`. +* `bwa/mergedLibrary/deepTools/plotFingerprint/` + * `*.plotFingerprint.pdf`, `*.plotFingerprint.qcmetrics.txt`, `*.plotFingerprint.raw.txt`: plotFingerprint output files. +* `bwa/mergedLibrary/deepTools/plotProfile/` + * `*.computeMatrix.mat.gz`, `*.computeMatrix.vals.mat.tab`, `*.plotProfile.pdf`, `*.plotProfile.tab`, `*.plotHeatmap.pdf`, `*.plotHeatmap.mat.tab`: plotProfile output files.
@@ -140,23 +141,22 @@ The results from deepTools plotProfile gives you a quick visualisation for the g ### Call peaks -[MACS2](https://github.com/taoliu/MACS) is one of the most popular peak-calling algorithms for ChIP-seq data. For ATAC-seq data we are also looking for genome-wide regions of enrichment but in this case without comparison to a standard control sample (e.g. input DNA). By default, the peaks are called with the MACS2 `--broad` parameter. If, however, you would like to call narrow peaks then please provide the `--narrow_peak` parameter when running the pipeline. See [MACS2 outputs](https://github.com/taoliu/MACS#output-files) for a description of the output files generated by MACS2. +[MACS2](https://github.com/taoliu/MACS) is one of the most popular peak-calling algorithms for ChIP-seq data. For ATAC-seq data we are also looking for genome-wide regions of enrichment but in this case without comparison to a standard control sample (e.g. input DNA). By default, the peaks are called with the MACS2 `--broad` parameter. If, however, you would like to call narrow peaks then please provide the `--narrow_peak` parameter when running the pipeline. See [MACS2 outputs](https://github.com/taoliu/MACS#output-files) for a description of the output files generated by MACS2. -[HOMER annotatePeaks.pl](http://homer.ucsd.edu/homer/ngs/annotation.html) is used to annotate the peaks relative to known genomic features. HOMER is able to use the `--gtf` annotation file which is provided to the pipeline. Please note that some of the output columns will be blank because the annotation is not provided using HOMER's in-built database format. However, the more important fields required for downstream analysis will be populated i.e. *Annotation*, *Distance to TSS* and *Nearest Promoter ID*. +[HOMER annotatePeaks.pl](http://homer.ucsd.edu/homer/ngs/annotation.html) is used to annotate the peaks relative to known genomic features. HOMER is able to use the `--gtf` annotation file which is provided to the pipeline. Please note that some of the output columns will be blank because the annotation is not provided using HOMER's in-built database format. However, the more important fields required for downstream analysis will be populated i.e. *Annotation*, *Distance to TSS* and *Nearest Promoter ID*. -Various QC plots per sample including number of peaks, fold-change distribution, [FRiP score](https://genome.cshlp.org/content/22/9/1813.full.pdf+html) and peak-to-gene feature annotation are also generated by the pipeline. Where possible these have been integrated into the MultiQC report. +Various QC plots per sample including number of peaks, fold-change distribution, [FRiP score](https://genome.cshlp.org/content/22/9/1813.full.pdf+html) and peak-to-gene feature annotation are also generated by the pipeline. Where possible these have been integrated into the MultiQC report.
Output files -* `bwa/mergedLibrary/macs//` - * MACS2 output files: `*.xls`, `*.broadPeak` or `*.narrowPeak`, `*.gappedPeak` and `*summits.bed`. - The files generated will depend on whether MACS2 has been run in *narrowPeak* or *broadPeak* mode. - * HOMER peak-to-gene annotation file: `*.annotatePeaks.txt`. -* `bwa/mergedLibrary/macs//qc/` - * QC plots for MACS2 peaks: `macs_peak.plots.pdf` - * QC plots for peak-to-gene feature annotation: `macs_annotatePeaks.plots.pdf` - * MultiQC custom-content files for FRiP score, peak count and peak-to-gene ratios: `*.FRiP_mqc.tsv`, `*.count_mqc.tsv` and `macs_annotatePeaks.summary_mqc.tsv` respectively. +* `bwa/mergedLibrary/macs//` + * `*.xls`, `*.broadPeak` or `*.narrowPeak`, `*.gappedPeak`, `*summits.bed`: MACS2 output files - the files generated will depend on whether MACS2 has been run in *narrowPeak* or *broadPeak* mode. + * `*.annotatePeaks.txt`: HOMER peak-to-gene annotation file. +* `bwa/mergedLibrary/macs//qc/` + * `macs_peak.plots.pdf`: QC plots for MACS2 peaks. + * `macs_annotatePeaks.plots.pdf`: QC plots for peak-to-gene feature annotation. + * `*.FRiP_mqc.tsv`, `*.count_mqc.tsv`, `macs_annotatePeaks.summary_mqc.tsv`: MultiQC custom-content files for FRiP score, peak count and peak-to-gene ratios. > **NB:** `` in the directory structure above corresponds to the type of peak that you have specified to call with MACS2 i.e. `broadPeak` or `narrowPeak`. If you so wish, you can call both narrow and broad peaks without redoing the preceding steps in the pipeline such as the alignment and filtering. For example, if you already have broad peaks then just add `--narrow_peak -resume` to the command you used to run the pipeline, and these will be called too! However, resuming the pipeline will only be possible if you have not deleted the `work/` directory generated by the pipeline. @@ -170,7 +170,7 @@ Various QC plots per sample including number of peaks, fold-change distribution, ### Create and quantify consensus set of peaks -In order to perform the differential binding analysis we need to be able to carry out the read quantification for the same intervals across **all** of the samples in the experiment. To this end, the individual peak-sets called per sample have to be merged together in order to create a consensus set of peaks. +In order to perform the differential accessibility analysis we need to be able to carry out the read quantification for the same intervals across **all** of the samples in the experiment. To this end, the individual peak-sets called per sample have to be merged together in order to create a consensus set of peaks. Using the consensus peaks it is possible to assess the degree of overlap between the peaks from a set of samples e.g. *Which consensus peaks contain peaks that are common/unique to a given set of samples?*. This may be useful for downstream filtering of peaks based on whether they are called in multiple replicates/conditions. Please note that it is possible for a consensus peak to contain multiple peaks from the same sample. Unfortunately, this is sample-dependent but the files generated by the pipeline do have columns that report such instances and allow you to factor them into any further analysis. @@ -181,47 +181,45 @@ The [featureCounts](http://bioinf.wehi.edu.au/featureCounts/) tool is used to co
Output files -* `bwa/mergedLibrary/macs//consensus/` - * Consensus peak-set across all samples in `*.bed` format. - * Consensus peak-set across all samples in `*.saf` format. Required by featureCounts for read quantification. - * `.featureCounts.txt` file for read counts across all samples relative to consensus peak-set. - * HOMER `*.annotatePeaks.txt` peak-to-gene annotation file for consensus peaks. - * Spreadsheet representation of consensus peak-set across samples **with** gene annotation columns: `*.boolean.annotatePeaks.txt`. - The columns from individual peak files are included in this file along with the ability to filter peaks based on their presence or absence in multiple replicates/conditions. - * Spreadsheet representation of consensus peak-set across samples **without** gene annotation columns: `*.boolean.txt`. - Same as file above but without annotation columns. - * [UpSetR](https://cran.r-project.org/web/packages/UpSetR/README.html) files to illustrate peak intersection: `*.boolean.intersect.plot.pdf` and `*.boolean.intersect.txt`. +* `bwa/mergedLibrary/macs//consensus/` + * `*.bed`: Consensus peak-set across all samples in BED format. + * `*.saf`: Consensus peak-set across all samples in SAF format. Required by featureCounts for read quantification. + * `*.featureCounts.txt`: Read counts across all samples relative to consensus peak-set. + * `*.annotatePeaks.txt`: HOMER peak-to-gene annotation file for consensus peaks. + * `*.boolean.annotatePeaks.txt`: Spreadsheet representation of consensus peak-set across samples **with** gene annotation columns. The columns from individual peak files are included in this file along with the ability to filter peaks based on their presence or absence in multiple replicates/conditions. + * `*.boolean.txt`: Spreadsheet representation of consensus peak-set across samples **without** gene annotation columns. Same as file above but without annotation columns. + * `*.boolean.intersect.plot.pdf`, `*.boolean.intersect.txt`: [UpSetR](https://cran.r-project.org/web/packages/UpSetR/README.html) files to illustrate peak intersection.
![R - UpSetR peak intersection plot](images/r_upsetr_intersect_plot.png) -![MultiQC - featureCounts consensus peak read assignment plot](images/mqc_featureCounts_assignment_plot.png) +![MultiQC - featureCounts consensus peak read assignment plot](images/mqc_featureCounts_assignment_plot.png) -### Read counting and differential binding analysis +### Read counting and differential accessibility analysis -[DESeq2](https://bioconductor.org/packages/release/bioc/vignettes/DESeq2/inst/doc/DESeq2.html) is more commonly used to perform differential expression analysis for RNA-seq datasets. However, it can also be used for ATAC-seq differential accessibility analysis, in which case you can imagine that instead of counts per gene for RNA-seq data we now have counts per accessible region. +[DESeq2](https://bioconductor.org/packages/release/bioc/vignettes/DESeq2/inst/doc/DESeq2.html) is more commonly used to perform differential expression analysis for RNA-seq datasets. However, it can also be used for ATAC-seq differential accessibility analysis, in which case you can imagine that instead of counts per gene for RNA-seq data we now have counts per accessible region. -This pipeline uses a standardised DESeq2 analysis script to get an idea of the reproducibility within the experiment, and to assess the overall differential binding. Please note that this will not suit every experimental design, and if there are other problems with the experiment then it may not work as well as expected. For larger experiments, it may be recommended to use the `vst` transformation instead of the default `rlog` option. You can do this by providing the `--deseq2_vst` parameter to the pipeline. See [DESeq2 docs](http://bioconductor.org/packages/devel/bioc/vignettes/DESeq2/inst/doc/DESeq2.html#data-transformations-and-visualization) for a more detailed explanation. +This pipeline uses a standardised DESeq2 analysis script to get an idea of the reproducibility within the experiment, and to assess the overall differential accessibility. Please note that this will not suit every experimental design, and if there are other problems with the experiment then it may not work as well as expected. For larger experiments, it may be recommended to use the `vst` transformation instead of the default `rlog` option. You can do this by providing the `--deseq2_vst` parameter to the pipeline. See [DESeq2 docs](http://bioconductor.org/packages/devel/bioc/vignettes/DESeq2/inst/doc/DESeq2.html#data-transformations-and-visualization) for a more detailed explanation. -By default, all possible pairwise comparisons across the groups from a particular antibody (as defined in [`design.csv`](usage.md#--design)) are performed. The DESeq2 results are generated by the pipeline in various ways. You can load up the results across all of the comparisons in a single spreadsheet, or individual folders will also be created that contain the results specific to a particular comparison. For the latter, additional files will also be generated where the intervals have been pre-filtered based on a couple of standard FDR thresholds. Please see [DESeq2 output](http://bioconductor.org/packages/release/bioc/vignettes/DESeq2/inst/doc/DESeq2.html#differential-expression-analysis) for a description of the columns generated by DESeq2. +By default, all possible pairwise comparisons across the groups are performed (as defined in [`design.csv`](usage.md#--design)). The DESeq2 results are generated by the pipeline in various ways. You can load up the results across all of the comparisons in a single spreadsheet, or individual folders will also be created that contain the results specific to a particular comparison. For the latter, additional files will also be generated where the intervals have been pre-filtered based on a couple of standard FDR thresholds. Please see [DESeq2 output](http://bioconductor.org/packages/release/bioc/vignettes/DESeq2/inst/doc/DESeq2.html#differential-expression-analysis) for a description of the columns generated by DESeq2.
Output files -* `bwa/mergedLibrary/macs//consensus//deseq2/` - * Differential binding `*.results.txt` spreadsheet containing results across all consensus peaks and all comparisons. - * `*.plots.pdf` file for PCA and hierarchical clustering. - * `*.log` file with information for number of differentially bound intervals at different FDR and fold-change thresholds for each comparison. - * `*.dds.rld.RData` file containing R `dds` and `rld` objects generated by DESeq2. - * `R_sessionInfo.log` file containing information about R, the OS and attached or loaded packages. -* `bwa/mergedLibrary/macs//consensus///` - * `*.results.txt` spreadsheet containing comparison-specific DESeq2 output for differential binding results across all peaks. - * Subset of above file for peaks that pass FDR <= 0.01 (`*FDR0.01.results.txt`) and FDR <= 0.05 (`*FDR0.05.results.txt`). - * BED files for peaks that pass FDR <= 0.01 (`*FDR0.01.results.bed`) and FDR <= 0.05 (`*FDR0.05.results.bed`). - * MA, Volcano, clustering and scatterplots at FDR <= 0.01 and FDR <= 0.05: `*deseq2.plots.pdf`. -* `bwa/mergedLibrary/macs//consensus//sizeFactors/` - Files containing DESeq2 sizeFactors per sample: `*.txt` and `*.RData`. +* `bwa/mergedLibrary/macs//consensus/deseq2/` + * `*.results.txt`: Spreadsheet containing differential accessibility results across all consensus peaks and all comparisons. + * `*.plots.pdf`: File containing PCA and hierarchical clustering plots. + * `*.log`: Log file with information for number of differentially accessible intervals at different FDR and fold-change thresholds for each comparison. + * `*.dds.rld.RData`: File containing R `dds` and `rld` objects generated by DESeq2. + * `R_sessionInfo.log`: File containing information about R, the OS and attached or loaded packages. +* `bwa/mergedLibrary/macs//consensus//` + * `*.results.txt`: Spreadsheet containing comparison-specific DESeq2 output for differential accessibility results across all peaks. + * `*FDR0.01.results.txt`, `*FDR0.05.results.txt`: Subset of above file for peaks that pass FDR <= 0.01 and FDR <= 0.05. + * `*FDR0.01.results.bed`, `*FDR0.05.results.bed`: BED files for peaks that pass FDR <= 0.01 and FDR <= 0.05. + * `*deseq2.plots.pdf`: MA, Volcano, clustering and scatterplots at FDR <= 0.01 and FDR <= 0.05. +* `bwa/mergedLibrary/macs//consensus/sizeFactors/` + * `*.txt`, `*.RData`: Files containing DESeq2 sizeFactors per sample.
@@ -240,20 +238,20 @@ By default, all possible pairwise comparisons across the groups from a particula
Output files -* `bwa/mergedLibrary/ataqv//` - `.json` files containing ATAC-seq specific metrics for each sample. -* `bwa/mergedLibrary/ataqv//html/` - Folder containing ataqv results aggregated across all samples for visualisation via an internet browser. +* `bwa/mergedLibrary/ataqv//` + * `.json`: JSON files containing ATAC-seq specific metrics for each sample. +* `bwa/mergedLibrary/ataqv//html/` + * Folder containing ataqv results aggregated across all samples for visualisation via an internet browser.
## Merged replicate-level analysis -The alignments associated with all of the replicates from the same experimental condition can also be merged. This can be useful to increase the coverage for peak-calling and for other analyses that require high sequencing depth such as [motif footprinting](https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3959825/). The analysis steps and directory structure for `bwa/mergedLibrary/` and `bwa/mergedReplicate/` are almost identical. +The alignments associated with all of the replicates from the same experimental condition can also be merged. This can be useful to increase the coverage for peak-calling and for other analyses that require high sequencing depth such as [motif footprinting](https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3959825/). The analysis steps and directory structure for `bwa/mergedLibrary/` and `bwa/mergedReplicate/` are almost identical. File names in the resulting directory (i.e. `bwa/mergedReplicate/`) will have the '`.mRp.`' suffix. -You can skip this portion of the analysis by specifying the `--skip_merge_replicates` parameter. +You can skip this portion of the analysis by specifying the `--skip_merge_replicates` parameter. > **NB:** Merged library-level alignments will be used for read counting relative to the consensus merged replicate-level peakset. This is the only way in which differential analysis can be performed at the merged replicate-level. @@ -261,19 +259,19 @@ You can skip this portion of the analysis by specifying the `--skip_merge_replic ### Present QC for the raw read, alignment, peak and differential accessibility results -[MultiQC](https://multiqc.info/docs/) is a visualisation tool that generates a single HTML report summarising all samples in your project. Most of the pipeline QC results are visualised in the report and further statistics are available within the report data directory. +[MultiQC](https://multiqc.info/docs/) is a visualisation tool that generates a single HTML report summarising all samples in your project. Most of the pipeline QC results are visualised in the report and further statistics are available within the report data directory. -Results generated by MultiQC collate pipeline QC from FastQC, TrimGalore, samtools flagstat, samtools idxstats, samtools stats, picard CollectMultipleMetrics, picard MarkDuplicates, Preseq, deepTools plotProfile, deepTools plotFingerprint and featureCounts. The default [`multiqc config file`](../assets/multiqc_config.yaml) also contains the provision for loading custom-content to report peak counts, FRiP scores, peak-to-gene annnotation proportions, sample-similarity heatmaps and PCA plots. +Results generated by MultiQC collate pipeline QC from FastQC, TrimGalore, samtools flagstat, samtools idxstats, samtools stats, picard CollectMultipleMetrics, picard MarkDuplicates, Preseq, deepTools plotProfile, deepTools plotFingerprint and featureCounts. The default [`multiqc config file`](../assets/multiqc_config.yaml) also contains the provision for loading custom-content to report peak counts, FRiP scores, peak-to-gene annnotation proportions, sample-similarity heatmaps and PCA plots. The pipeline has special steps which also allow the software versions to be reported in the MultiQC output for future traceability. For more information about how to use MultiQC reports, see .
Output files -* `multiqc//` - * `multiqc_report.html` - a standalone HTML file that can be viewed in your web browser. - * `multiqc_data/` - directory containing parsed statistics from the different tools used in the pipeline. - * `multiqc_plots/` - directory containing static images from the report in various formats. +* `multiqc//` + * `multiqc_report.html`: A standalone HTML file that can be viewed in your web browser. + * `multiqc_data/`: Directory containing parsed statistics from the different tools used in the pipeline. + * `multiqc_plots/`: Directory containing static images from the report in various formats.
@@ -292,9 +290,9 @@ Once installed, open IGV, go to `File > Open Session` and select the `igv_sessio
Output files -* `igv//` - * `igv_session.xml` file. - * `igv_files.txt` file containing a listing of the files used to create the IGV session, and their allocated colours. +* `igv//` + * `igv_session.xml`: Session file that can be directly loaded into IGV. + * `igv_files.txt`: File containing a listing of the files used to create the IGV session.
@@ -309,10 +307,10 @@ Reference genome-specific files can be useful to keep for the downstream process
Output files -* `genome/` - A number of genome-specific files are generated by the pipeline in order to aid in the filtering of the data, and because they are required by standard tools such as BEDTools. These can be found in this directory along with the genome fasta file which is required by IGV. If using a genome from AWS iGenomes and if it exists a `README.txt` file containing information about the annotation version will also be saved in this directory. -* `genome/BWAIndex/` - If the `--save_reference` parameter is provided then the alignment indices generated by the pipeline will be saved in this directory. This can be quite a time-consuming process so it permits their reuse for future runs of the pipeline or for other purposes. +* `genome/` + * A number of genome-specific files are generated by the pipeline in order to aid in the filtering of the data, and because they are required by standard tools such as BEDTools. These can be found in this directory along with the genome fasta file which is required by IGV. If using a genome from AWS iGenomes and if it exists a `README.txt` file containing information about the annotation version will also be saved in this directory. +* `genome/BWAIndex/` + * If the `--save_reference` parameter is provided then the alignment indices generated by the pipeline will be saved in this directory. This can be quite a time-consuming process so it permits their reuse for future runs of the pipeline or for other purposes.
@@ -323,11 +321,10 @@ Reference genome-specific files can be useful to keep for the downstream process
Output files -* `pipeline_info/` - * Reports generated by the pipeline - `pipeline_report.html`, `pipeline_report.txt` and `software_versions.csv`. - * Reports generated by Nextflow - `execution_report.html`, `execution_timeline.html`, `execution_trace.txt` and `pipeline_dag.svg`. - * Reformatted design files used as input to the pipeline - `design_reads.csv` and `design_controls.csv`. -* `Documentation/` - Documentation for interpretation of results in HTML format - `results_description.html`. +* `pipeline_info/` + * `pipeline_report.html`, `pipeline_report.txt`, `software_versions.csv`: Reports generated by the pipeline. + * `execution_report.html`, `execution_timeline.html`, `execution_trace.txt`, `pipeline_dag.svg`: Reports generated by Nextflow. + * `design_reads.csv`: Reformatted design files used as input to the pipeline. + * `results_description.html`: Documentation for interpretation of results in HTML format.
From 9262aa0544b67a5443cdc21adb44718d31103232 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 30 Jun 2020 16:15:22 +0100 Subject: [PATCH 207/226] Change output dir --- main.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/main.nf b/main.nf index 1f040e2a..291de879 100644 --- a/main.nf +++ b/main.nf @@ -2047,7 +2047,7 @@ process MULTIQC { * STEP 11: Output description HTML */ process output_documentation { - publishDir "${params.outdir}/Documentation", mode: params.publish_dir_mode + publishDir "${params.outdir}/pipeline_info", mode: params.publish_dir_mode input: path output_docs from ch_output_docs From 7c9d1f7971a7beae56735342f555f1ef4e56a5e4 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 30 Jun 2020 17:54:51 +0100 Subject: [PATCH 208/226] Update packages in environment --- .github/markdownlint.yml | 2 +- CHANGELOG.md | 10 +++++----- environment.yml | 8 ++++---- 3 files changed, 10 insertions(+), 10 deletions(-) diff --git a/.github/markdownlint.yml b/.github/markdownlint.yml index 9aae63a0..5056c7fa 100644 --- a/.github/markdownlint.yml +++ b/.github/markdownlint.yml @@ -4,6 +4,6 @@ line-length: false no-duplicate-header: siblings_only: true MD033: - allowed_elements: [details, summary] + allowed_elements: [details, summary, p, img] MD007: indent: 4 diff --git a/CHANGELOG.md b/CHANGELOG.md index 7a79a3c8..21f5a30d 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -30,11 +30,11 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * [#73](https://github.com/nf-core/atacseq/issues/73) - macs_annotatePeaks.mLb.clN.summary.txt file is not created * [#86](https://github.com/nf-core/atacseq/issues/86) - bug in the plot_homer_annotatepeaks.r script * [#102](https://github.com/nf-core/atacseq/issues/102) - Incorrect Group ID assigned by featurecounts_deseq2.r +* [#110](https://github.com/nf-core/atacseq/pull/110) - updated AWS test GitHub actions * [nf-core/chipseq#118](https://github.com/nf-core/chipseq/issues/118) - Running on with SGE * [nf-core/chipseq#132](https://github.com/nf-core/chipseq/issues/132) - BigWig Error: sort: cannot create temporary file in '': Read-only file system * [nf-core/chipseq#154](https://github.com/nf-core/chipseq/issues/154) - computeMatrix.val.mat.gz files not zipped * Make executables in `bin/` compatible with Python 3 -* [#110](https://github.com/nf-core/atacseq/pull/110) - updated AWS test GitHub actions ### `Dependencies` @@ -49,16 +49,16 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * Update gawk `4.2.1` -> `5.1.0` * Update r-base `3.4.1` -> `3.6.2` * Update r-optparse `1.6.0` -> `1.6.6` -* Update r-ggplot2 `3.1.0` -> `3.3.0` +* Update r-ggplot2 `3.1.0` -> `3.3.2` * Update r-pheatmap `1.0.10` -> `1.0.12` * Update r-lattice `0.20_35` -> `0.20_41` * Update r-upsetr `1.3.3` -> `1.4.0` * Update r-scales `1.0.0` -> `1.1.1` -* Update r-xfun `0.3` -> `0.14` +* Update r-xfun `0.3` -> `0.15` * Update fastqc `0.11.8` -> `0.11.9` * Update trim-galore `0.5.0` -> `0.6.5` * Update samtools `1.9` -> `1.10` -* Update picard `2.19.0` -> `2.22.8` +* Update picard `2.19.0` -> `2.23.1` * Update pysam `0.15.2` -> `0.15.3` * Update bedtools `2.27.1` -> `2.29.2` * Update ucsc-bedgraphtobigwig `377` -> `357` @@ -66,7 +66,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * Update macs2 `2.1.2` -> `2.2.7.1` * Update homer `4.9.1` -> `4.11` * Update ataqv `1.0.0` -> `1.1.1` -* Update subread `1.6.4` -> `2.0.0` +* Update subread `1.6.4` -> `2.0.1` * Update multiqc `1.7` -> `1.8` * Update bioconductor-deseq2 `1.20.0` -> `1.26.0` * Update bioconductor-vsn `3.46.0` -> `3.54.0` diff --git a/environment.yml b/environment.yml index 66ce39c7..cadc2140 100755 --- a/environment.yml +++ b/environment.yml @@ -15,13 +15,13 @@ dependencies: - conda-forge::r-optparse=1.6.6 - conda-forge::r-rcolorbrewer=1.1_2 - conda-forge::r-reshape2=1.4.4 - - conda-forge::r-ggplot2=3.3.0 + - conda-forge::r-ggplot2=3.3.2 - conda-forge::r-tidyr=1.1.0 - conda-forge::r-scales=1.1.1 - conda-forge::r-pheatmap=1.0.12 - conda-forge::r-lattice=0.20_41 - conda-forge::r-upsetr=1.4.0 - - conda-forge::r-xfun=0.14 + - conda-forge::r-xfun=0.15 - conda-forge::gawk=5.1.0 - conda-forge::pigz=2.3.4 ## Required for TrimGalore multi-threading @@ -30,7 +30,7 @@ dependencies: - bioconda::trim-galore=0.6.5 - bioconda::bwa=0.7.17 - bioconda::samtools=1.10 - - bioconda::picard=2.22.8 + - bioconda::picard=2.23.1 - bioconda::bamtools=2.5.1 - bioconda::pysam=0.15.3 - bioconda::bedtools=2.29.2 @@ -39,7 +39,7 @@ dependencies: - bioconda::macs2=2.2.7.1 - bioconda::homer=4.11 - bioconda::ataqv=1.1.1 - - bioconda::subread=2.0.0 + - bioconda::subread=2.0.1 - bioconda::preseq=2.0.3 - bioconda::multiqc=1.9 - bioconda::bioconductor-biocparallel=1.20.0 From 5ea601efe28c01bde203fda78b145e4bff528643 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 30 Jun 2020 18:09:28 +0100 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Output files @@ -28,9 +26,9 @@ The initial QC and alignments are performed at the library-level e.g. if the sam
-### Adapter trimming +[FastQC](http://www.bioinformatics.babraham.ac.uk/projects/fastqc/Help/) gives general quality metrics about your reads. It provides information about the quality score distribution across your reads, the per base sequence content (%A/C/G/T). You get information about adapter contamination and other overrepresented sequences. -[Trim Galore!](https://www.bioinformatics.babraham.ac.uk/projects/trim_galore/) is a wrapper tool around Cutadapt and FastQC to consistently apply quality and adapter trimming to FastQ files. By default, Trim Galore! will automatically detect and trim the appropriate adapter sequence. For most ATAC-seq datasets this will be the Nextera adapter sequence 'CTGTCTCTTATA'. See [`usage.md`](usage.md) for more details about the trimming options. +### Adapter trimming
Output files @@ -46,12 +44,12 @@ The initial QC and alignments are performed at the library-level e.g. if the sam
+[Trim Galore!](https://www.bioinformatics.babraham.ac.uk/projects/trim_galore/) is a wrapper tool around Cutadapt and FastQC to consistently apply quality and adapter trimming to FastQ files. By default, Trim Galore! will automatically detect and trim the appropriate adapter sequence. For most ATAC-seq datasets this will be the Nextera adapter sequence 'CTGTCTCTTATA'. See [`usage.md`](usage.md) for more details about the trimming options. + ![MultiQC - Cutadapt trimmed sequence plot](images/mqc_cutadapt_plot.png) ### Alignment -Adapter-trimmed reads are mapped to the reference assembly using [BWA](http://bio-bwa.sourceforge.net/bwa.shtml). A genome index is required to run BWA so if this is not provided explicitly using the `--bwa_index` parameter then it will be created automatically from the genome fasta input. The index creation process can take a while for larger genomes so it is possible to use the `--save_reference` parameter to save the indices for future pipeline runs, reducing processing times. -
Output files @@ -64,6 +62,8 @@ Adapter-trimmed reads are mapped to the reference assembly using [BWA](http://bi
+Adapter-trimmed reads are mapped to the reference assembly using [BWA](http://bio-bwa.sourceforge.net/bwa.shtml). A genome index is required to run BWA so if this is not provided explicitly using the `--bwa_index` parameter then it will be created automatically from the genome fasta input. The index creation process can take a while for larger genomes so it is possible to use the `--save_reference` parameter to save the indices for future pipeline runs, reducing processing times. + ![MultiQC - SAMtools stats plot](images/mqc_samtools_stats_plot.png) ![MultiQC - SAMtools idxstats plot](images/mqc_samtools_idxstats_plot.png) @@ -74,14 +74,6 @@ The library-level alignments associated with the same sample are merged and subs ### Alignment merging, duplicate marking, filtering and QC -[Picard MergeSamFiles and MarkDuplicates](https://broadinstitute.github.io/picard/command-line-overview.html) are used in combination to merge the alignments, and for the marking of duplicates, respectively. If you only have one library for any given replicate then the merging step isnt carried out because the library-level and merged library-level BAM files will be exactly the same. - -Read duplicate marking is carried out using the Picard MarkDuplicates command. Duplicate reads are generally removed from the aligned reads to mitigate for fragments in the library that may have been sequenced more than once due to PCR biases. There is an option to keep duplicate reads with the `--keep_dups` parameter but its generally recommended to remove them to avoid the wrong interpretation of the results. A similar option has been provided to keep reads that are multi-mapped - `--keep_multi_map`. Other steps have been incorporated into the pipeline to filter the resulting alignments - see [`main README.md`](../README.md) for a more comprehensive listing, and the tools used at each step. A selection of alignment-based QC metrics generated by Picard CollectMultipleMetrics and MarkDuplicates will be included in the MultiQC report. - -Certain cell types and tissues yield an enormous fraction (typically 20–80%) of unusable sequences of mitochondrial origin. This is a known problem that is specific to ATAC-seq library preps - see [Montefiori et al. 2017](https://www.nature.com/articles/s41598-017-02547-w). There is an option to keep these reads using the `--keep_mito` parameter but its generally recommended to remove these in order to get a more reliable assessment of the duplication rate from the rest of the genome, and to avoid any biases in the downstream analyses. - -The [Preseq](http://smithlabresearch.org/software/preseq/) package is aimed at predicting and estimating the complexity of a genomic sequencing library, equivalent to predicting and estimating the number of redundant reads from a given sequencing depth and how many will be expected from additional sequencing using an initial sequencing experiment. The estimates can then be used to examine the utility of further sequencing, optimize the sequencing depth, or to screen multiple libraries to avoid low complexity samples. The dashed line shows a perfectly complex library where total reads = unique reads. Note that these are predictive numbers only, not absolute. The MultiQC plot can sometimes give extreme sequencing depth on the X axis - click and drag from the left side of the plot to zoom in on more realistic numbers. -
Output files @@ -101,16 +93,22 @@ The [Preseq](http://smithlabresearch.org/software/preseq/) package is aimed at p
+[Picard MergeSamFiles and MarkDuplicates](https://broadinstitute.github.io/picard/command-line-overview.html) are used in combination to merge the alignments, and for the marking of duplicates, respectively. If you only have one library for any given replicate then the merging step isnt carried out because the library-level and merged library-level BAM files will be exactly the same. + ![MultiQC - Picard deduplication stats plot](images/mqc_picard_deduplication_plot.png) +Read duplicate marking is carried out using the Picard MarkDuplicates command. Duplicate reads are generally removed from the aligned reads to mitigate for fragments in the library that may have been sequenced more than once due to PCR biases. There is an option to keep duplicate reads with the `--keep_dups` parameter but its generally recommended to remove them to avoid the wrong interpretation of the results. A similar option has been provided to keep reads that are multi-mapped - `--keep_multi_map`. Other steps have been incorporated into the pipeline to filter the resulting alignments - see [`main README.md`](../README.md) for a more comprehensive listing, and the tools used at each step. A selection of alignment-based QC metrics generated by Picard CollectMultipleMetrics and MarkDuplicates will be included in the MultiQC report. + ![MultiQC - Picard insert size plot](images/mqc_picard_insert_size_plot.png) +Certain cell types and tissues yield an enormous fraction (typically 20–80%) of unusable sequences of mitochondrial origin. This is a known problem that is specific to ATAC-seq library preps - see [Montefiori et al. 2017](https://www.nature.com/articles/s41598-017-02547-w). There is an option to keep these reads using the `--keep_mito` parameter but its generally recommended to remove these in order to get a more reliable assessment of the duplication rate from the rest of the genome, and to avoid any biases in the downstream analyses. + +The [Preseq](http://smithlabresearch.org/software/preseq/) package is aimed at predicting and estimating the complexity of a genomic sequencing library, equivalent to predicting and estimating the number of redundant reads from a given sequencing depth and how many will be expected from additional sequencing using an initial sequencing experiment. The estimates can then be used to examine the utility of further sequencing, optimize the sequencing depth, or to screen multiple libraries to avoid low complexity samples. The dashed line shows a perfectly complex library where total reads = unique reads. Note that these are predictive numbers only, not absolute. The MultiQC plot can sometimes give extreme sequencing depth on the X axis - click and drag from the left side of the plot to zoom in on more realistic numbers. + ![MultiQC - Preseq library complexity plot](images/mqc_preseq_plot.png) ### Normalised bigWig files -The [bigWig](https://genome.ucsc.edu/goldenpath/help/bigWig.html) format is in an indexed binary format useful for displaying dense, continuous data in Genome Browsers such as the [UCSC](https://genome.ucsc.edu/cgi-bin/hgTracks) and [IGV](http://software.broadinstitute.org/software/igv/). This mitigates the need to load the much larger BAM files for data visualisation purposes which will be slower and result in memory issues. The coverage values represented in the bigWig file can also be normalised in order to be able to compare the coverage across multiple samples - this is not possible with BAM files. The bigWig format is also supported by various bioinformatics software for downstream processing such as meta-profile plotting. -
Output files @@ -119,11 +117,9 @@ The [bigWig](https://genome.ucsc.edu/goldenpath/help/bigWig.html) format is in a
-### Coverage QC - -[deepTools](https://deeptools.readthedocs.io/en/develop/content/list_of_tools.html) plotFingerprint is a useful QC for ATAC-seq data in order to see the relative enrichment of the samples in the experiment on a genome-wide basis (see [plotFingerprint docs](https://deeptools.readthedocs.io/en/develop/content/tools/plotFingerprint.html)). +The [bigWig](https://genome.ucsc.edu/goldenpath/help/bigWig.html) format is in an indexed binary format useful for displaying dense, continuous data in Genome Browsers such as the [UCSC](https://genome.ucsc.edu/cgi-bin/hgTracks) and [IGV](http://software.broadinstitute.org/software/igv/). This mitigates the need to load the much larger BAM files for data visualisation purposes which will be slower and result in memory issues. The coverage values represented in the bigWig file can also be normalised in order to be able to compare the coverage across multiple samples - this is not possible with BAM files. The bigWig format is also supported by various bioinformatics software for downstream processing such as meta-profile plotting. -The results from deepTools plotProfile gives you a quick visualisation for the genome-wide enrichment of your samples at the TSS, and across the gene body. During the downstream analysis, you may want to refine the features/genes used to generate these plots in order to see a more specific condition-related effect. +### Coverage QC
Output files @@ -135,18 +131,16 @@ The results from deepTools plotProfile gives you a quick visualisation for the g
+[deepTools](https://deeptools.readthedocs.io/en/develop/content/list_of_tools.html) plotFingerprint is a useful QC for ATAC-seq data in order to see the relative enrichment of the samples in the experiment on a genome-wide basis (see [plotFingerprint docs](https://deeptools.readthedocs.io/en/develop/content/tools/plotFingerprint.html)). + ![MultiQC - deepTools plotFingerprint plot](images/mqc_deeptools_plotFingerprint_plot.png) +The results from deepTools plotProfile gives you a quick visualisation for the genome-wide enrichment of your samples at the TSS, and across the gene body. During the downstream analysis, you may want to refine the features/genes used to generate these plots in order to see a more specific condition-related effect. + ![MultiQC - deepTools plotProfile plot](images/mqc_deeptools_plotProfile_plot.png) ### Call peaks -[MACS2](https://github.com/taoliu/MACS) is one of the most popular peak-calling algorithms for ChIP-seq data. For ATAC-seq data we are also looking for genome-wide regions of enrichment but in this case without comparison to a standard control sample (e.g. input DNA). By default, the peaks are called with the MACS2 `--broad` parameter. If, however, you would like to call narrow peaks then please provide the `--narrow_peak` parameter when running the pipeline. See [MACS2 outputs](https://github.com/taoliu/MACS#output-files) for a description of the output files generated by MACS2. - -[HOMER annotatePeaks.pl](http://homer.ucsd.edu/homer/ngs/annotation.html) is used to annotate the peaks relative to known genomic features. HOMER is able to use the `--gtf` annotation file which is provided to the pipeline. Please note that some of the output columns will be blank because the annotation is not provided using HOMER's in-built database format. However, the more important fields required for downstream analysis will be populated i.e. *Annotation*, *Distance to TSS* and *Nearest Promoter ID*. - -Various QC plots per sample including number of peaks, fold-change distribution, [FRiP score](https://genome.cshlp.org/content/22/9/1813.full.pdf+html) and peak-to-gene feature annotation are also generated by the pipeline. Where possible these have been integrated into the MultiQC report. -
Output files @@ -162,22 +156,20 @@ Various QC plots per sample including number of peaks, fold-change distribution,
+[MACS2](https://github.com/taoliu/MACS) is one of the most popular peak-calling algorithms for ChIP-seq data. For ATAC-seq data we are also looking for genome-wide regions of enrichment but in this case without comparison to a standard control sample (e.g. input DNA). By default, the peaks are called with the MACS2 `--broad` parameter. If, however, you would like to call narrow peaks then please provide the `--narrow_peak` parameter when running the pipeline. See [MACS2 outputs](https://github.com/taoliu/MACS#output-files) for a description of the output files generated by MACS2. + ![MultiQC - MACS2 total peak count plot](images/mqc_macs2_peak_count_plot.png) +[HOMER annotatePeaks.pl](http://homer.ucsd.edu/homer/ngs/annotation.html) is used to annotate the peaks relative to known genomic features. HOMER is able to use the `--gtf` annotation file which is provided to the pipeline. Please note that some of the output columns will be blank because the annotation is not provided using HOMER's in-built database format. However, the more important fields required for downstream analysis will be populated i.e. *Annotation*, *Distance to TSS* and *Nearest Promoter ID*. + ![MultiQC - HOMER annotatePeaks peak-to-gene feature ratio plot](images/mqc_annotatePeaks_feature_percentage_plot.png) +Various QC plots per sample including number of peaks, fold-change distribution, [FRiP score](https://genome.cshlp.org/content/22/9/1813.full.pdf+html) and peak-to-gene feature annotation are also generated by the pipeline. Where possible these have been integrated into the MultiQC report. + ![MultiQC - MACS2 peaks FRiP score plot](images/mqc_frip_score_plot.png) ### Create and quantify consensus set of peaks -In order to perform the differential accessibility analysis we need to be able to carry out the read quantification for the same intervals across **all** of the samples in the experiment. To this end, the individual peak-sets called per sample have to be merged together in order to create a consensus set of peaks. - -Using the consensus peaks it is possible to assess the degree of overlap between the peaks from a set of samples e.g. *Which consensus peaks contain peaks that are common/unique to a given set of samples?*. This may be useful for downstream filtering of peaks based on whether they are called in multiple replicates/conditions. Please note that it is possible for a consensus peak to contain multiple peaks from the same sample. Unfortunately, this is sample-dependent but the files generated by the pipeline do have columns that report such instances and allow you to factor them into any further analysis. - -By default, the peak-sets are not filtered, therefore, the consensus peaks will be generated across the union set of peaks from all samples. However, you can increment the `--min_reps_consensus` parameter appropriately if you are confident you have good reproducibility amongst your replicates to create a "reproducible" set of consensus of peaks. In future iterations of the pipeline more formal analyses such as [IDR](https://projecteuclid.org/euclid.aoas/1318514284) may be implemented to obtain reproducible and high confidence peak-sets with which to perform this sort of analysis. - -The [featureCounts](http://bioinf.wehi.edu.au/featureCounts/) tool is used to count the number of reads relative to the consensus peak-set across all of the samples. This essentially generates a file containing a matrix where the rows represent the consensus intervals, the columns represent all of the samples in the experiment, and the values represent the raw read counts. -
Output files @@ -192,17 +184,19 @@ The [featureCounts](http://bioinf.wehi.edu.au/featureCounts/) tool is used to co
-![R - UpSetR peak intersection plot](images/r_upsetr_intersect_plot.png) +In order to perform the differential accessibility analysis we need to be able to carry out the read quantification for the same intervals across **all** of the samples in the experiment. To this end, the individual peak-sets called per sample have to be merged together in order to create a consensus set of peaks. -![MultiQC - featureCounts consensus peak read assignment plot](images/mqc_featureCounts_assignment_plot.png) +Using the consensus peaks it is possible to assess the degree of overlap between the peaks from a set of samples e.g. *Which consensus peaks contain peaks that are common/unique to a given set of samples?*. This may be useful for downstream filtering of peaks based on whether they are called in multiple replicates/conditions. Please note that it is possible for a consensus peak to contain multiple peaks from the same sample. Unfortunately, this is sample-dependent but the files generated by the pipeline do have columns that report such instances and allow you to factor them into any further analysis. -### Read counting and differential accessibility analysis +![R - UpSetR peak intersection plot](images/r_upsetr_intersect_plot.png) -[DESeq2](https://bioconductor.org/packages/release/bioc/vignettes/DESeq2/inst/doc/DESeq2.html) is more commonly used to perform differential expression analysis for RNA-seq datasets. However, it can also be used for ATAC-seq differential accessibility analysis, in which case you can imagine that instead of counts per gene for RNA-seq data we now have counts per accessible region. +By default, the peak-sets are not filtered, therefore, the consensus peaks will be generated across the union set of peaks from all samples. However, you can increment the `--min_reps_consensus` parameter appropriately if you are confident you have good reproducibility amongst your replicates to create a "reproducible" set of consensus of peaks. In future iterations of the pipeline more formal analyses such as [IDR](https://projecteuclid.org/euclid.aoas/1318514284) may be implemented to obtain reproducible and high confidence peak-sets with which to perform this sort of analysis. -This pipeline uses a standardised DESeq2 analysis script to get an idea of the reproducibility within the experiment, and to assess the overall differential accessibility. Please note that this will not suit every experimental design, and if there are other problems with the experiment then it may not work as well as expected. For larger experiments, it may be recommended to use the `vst` transformation instead of the default `rlog` option. You can do this by providing the `--deseq2_vst` parameter to the pipeline. See [DESeq2 docs](http://bioconductor.org/packages/devel/bioc/vignettes/DESeq2/inst/doc/DESeq2.html#data-transformations-and-visualization) for a more detailed explanation. +The [featureCounts](http://bioinf.wehi.edu.au/featureCounts/) tool is used to count the number of reads relative to the consensus peak-set across all of the samples. This essentially generates a file containing a matrix where the rows represent the consensus intervals, the columns represent all of the samples in the experiment, and the values represent the raw read counts. -By default, all possible pairwise comparisons across the groups are performed (as defined in [`design.csv`](usage.md#--design)). The DESeq2 results are generated by the pipeline in various ways. You can load up the results across all of the comparisons in a single spreadsheet, or individual folders will also be created that contain the results specific to a particular comparison. For the latter, additional files will also be generated where the intervals have been pre-filtered based on a couple of standard FDR thresholds. Please see [DESeq2 output](http://bioconductor.org/packages/release/bioc/vignettes/DESeq2/inst/doc/DESeq2.html#differential-expression-analysis) for a description of the columns generated by DESeq2. +![MultiQC - featureCounts consensus peak read assignment plot](images/mqc_featureCounts_assignment_plot.png) + +### Read counting and differential accessibility analysis
Output files @@ -223,18 +217,26 @@ By default, all possible pairwise comparisons across the groups are performed (a
+[DESeq2](https://bioconductor.org/packages/release/bioc/vignettes/DESeq2/inst/doc/DESeq2.html) is more commonly used to perform differential expression analysis for RNA-seq datasets. However, it can also be used for ATAC-seq differential accessibility analysis, in which case you can imagine that instead of counts per gene for RNA-seq data we now have counts per accessible region. + +This pipeline uses a standardised DESeq2 analysis script to get an idea of the reproducibility within the experiment, and to assess the overall differential accessibility. Please note that this will not suit every experimental design, and if there are other problems with the experiment then it may not work as well as expected. For larger experiments, it may be recommended to use the `vst` transformation instead of the default `rlog` option. You can do this by providing the `--deseq2_vst` parameter to the pipeline. See [DESeq2 docs](http://bioconductor.org/packages/devel/bioc/vignettes/DESeq2/inst/doc/DESeq2.html#data-transformations-and-visualization) for a more detailed explanation. + ![MultiQC - DESeq2 PCA plot](images/mqc_deseq2_pca_plot.png) ![MultiQC - DESeq2 sample similarity plot](images/mqc_deseq2_sample_similarity_plot.png) -![R - DESeq2 MA plot](images/r_deseq2_ma_plot.png) +By default, all possible pairwise comparisons across the groups are performed (as defined in [`design.csv`](usage.md#--design)). The DESeq2 results are generated by the pipeline in various ways. You can load up the results across all of the comparisons in a single spreadsheet, or individual folders will also be created that contain the results specific to a particular comparison. For the latter, additional files will also be generated where the intervals have been pre-filtered based on a couple of standard FDR thresholds. Please see [DESeq2 output](http://bioconductor.org/packages/release/bioc/vignettes/DESeq2/inst/doc/DESeq2.html#differential-expression-analysis) for a description of the columns generated by DESeq2. + +

+ R - DESeq2 MA plot +

-![R - DESeq2 Volcano plot](images/r_deseq2_volcano_plot.png) +

+ R - DESeq2 Volcano plot +

### ataqv -[ataqv](https://parkerlab.github.io/ataqv/) is a toolkit for measuring and comparing ATAC-seq results. It was written to help understand how well ATAC-seq assays have worked, and to make it easier to spot differences that might be caused by library prep or sequencing. Please see [ataqv homepage](https://parkerlab.github.io/ataqv/) for documentation and an example report. -
Output files @@ -245,6 +247,8 @@ By default, all possible pairwise comparisons across the groups are performed (a
+[ataqv](https://parkerlab.github.io/ataqv/) is a toolkit for measuring and comparing ATAC-seq results. It was written to help understand how well ATAC-seq assays have worked, and to make it easier to spot differences that might be caused by library prep or sequencing. Please see [ataqv homepage](https://parkerlab.github.io/ataqv/) for documentation and an example report. + ## Merged replicate-level analysis The alignments associated with all of the replicates from the same experimental condition can also be merged. This can be useful to increase the coverage for peak-calling and for other analyses that require high sequencing depth such as [motif footprinting](https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3959825/). The analysis steps and directory structure for `bwa/mergedLibrary/` and `bwa/mergedReplicate/` are almost identical. @@ -259,12 +263,6 @@ You can skip this portion of the analysis by specifying the `--skip_merge_replic ### Present QC for the raw read, alignment, peak and differential accessibility results -[MultiQC](https://multiqc.info/docs/) is a visualisation tool that generates a single HTML report summarising all samples in your project. Most of the pipeline QC results are visualised in the report and further statistics are available within the report data directory. - -Results generated by MultiQC collate pipeline QC from FastQC, TrimGalore, samtools flagstat, samtools idxstats, samtools stats, picard CollectMultipleMetrics, picard MarkDuplicates, Preseq, deepTools plotProfile, deepTools plotFingerprint and featureCounts. The default [`multiqc config file`](../assets/multiqc_config.yaml) also contains the provision for loading custom-content to report peak counts, FRiP scores, peak-to-gene annnotation proportions, sample-similarity heatmaps and PCA plots. - -The pipeline has special steps which also allow the software versions to be reported in the MultiQC output for future traceability. For more information about how to use MultiQC reports, see . -
Output files @@ -275,17 +273,13 @@ The pipeline has special steps which also allow the software versions to be repo
-### Create IGV session file - -An [IGV](https://software.broadinstitute.org/software/igv/UserGuide) session file will be created at the end of the pipeline containing the normalised bigWig tracks, per-sample peaks, consensus peaks and differential sites. This avoids having to load all of the data individually into IGV for visualisation. - -The genome fasta file required for the IGV session will be the same as the one that was provided to the pipeline. This will be copied into `genome/` to overcome any loading issues. If you prefer to use another path or an in-built genome provided by IGV just change the `genome` entry in the second-line of the session file. +[MultiQC](https://multiqc.info/docs/) is a visualisation tool that generates a single HTML report summarising all samples in your project. Most of the pipeline QC results are visualised in the report and further statistics are available within the report data directory. -The file paths in the IGV session file will only work if the results are kept in the same place on your storage. If the results are moved or for example, if you prefer to load the data over the web then just replace the file paths with others that are more appropriate. +Results generated by MultiQC collate pipeline QC from FastQC, TrimGalore, samtools flagstat, samtools idxstats, samtools stats, picard CollectMultipleMetrics, picard MarkDuplicates, Preseq, deepTools plotProfile, deepTools plotFingerprint and featureCounts. The default [`multiqc config file`](../assets/multiqc_config.yaml) also contains the provision for loading custom-content to report peak counts, FRiP scores, peak-to-gene annnotation proportions, sample-similarity heatmaps and PCA plots. -Once installed, open IGV, go to `File > Open Session` and select the `igv_session.xml` file for loading. +The pipeline has special steps which also allow the software versions to be reported in the MultiQC output for future traceability. For more information about how to use MultiQC reports, see . -> **NB:** If you are not using an in-built genome provided by IGV you will need to load the annotation yourself e.g. in .gtf and/or .bed format. +### Create IGV session file
Output files @@ -296,14 +290,22 @@ Once installed, open IGV, go to `File > Open Session` and select the `igv_sessio
+An [IGV](https://software.broadinstitute.org/software/igv/UserGuide) session file will be created at the end of the pipeline containing the normalised bigWig tracks, per-sample peaks, consensus peaks and differential sites. This avoids having to load all of the data individually into IGV for visualisation. + +The genome fasta file required for the IGV session will be the same as the one that was provided to the pipeline. This will be copied into `genome/` to overcome any loading issues. If you prefer to use another path or an in-built genome provided by IGV just change the `genome` entry in the second-line of the session file. + +The file paths in the IGV session file will only work if the results are kept in the same place on your storage. If the results are moved or for example, if you prefer to load the data over the web then just replace the file paths with others that are more appropriate. + +Once installed, open IGV, go to `File > Open Session` and select the `igv_session.xml` file for loading. + ![IGV screenshot](images/igv_screenshot.png) +> **NB:** If you are not using an in-built genome provided by IGV you will need to load the annotation yourself e.g. in .gtf and/or .bed format. + ## Other results ### Reference genome files -Reference genome-specific files can be useful to keep for the downstream processing of the results. -
Output files @@ -314,9 +316,9 @@ Reference genome-specific files can be useful to keep for the downstream process
-### Pipeline information +Reference genome-specific files can be useful to keep for the downstream processing of the results. -[Nextflow](https://www.nextflow.io/docs/latest/tracing.html) provides excellent functionality for generating various reports relevant to the running and execution of the pipeline. This will allow you to trouble-shoot errors with the running of the pipeline, and also provide you with other information such as launch commands, run times and resource usage. +### Pipeline information
Output files @@ -328,3 +330,5 @@ Reference genome-specific files can be useful to keep for the downstream process * `results_description.html`: Documentation for interpretation of results in HTML format.
+ +[Nextflow](https://www.nextflow.io/docs/latest/tracing.html) provides excellent functionality for generating various reports relevant to the running and execution of the pipeline. This will allow you to trouble-shoot errors with the running of the pipeline, and also provide you with other information such as launch commands, run times and resource usage. From 1dc03f3026e2a88f70a7280a66c065f30f2677a6 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 30 Jun 2020 18:43:09 +0100 Subject: [PATCH 210/226] Update docs images --- ..._annotatePeaks_feature_percentage_plot.png | Bin 73104 -> 24552 bytes docs/images/mqc_cutadapt_plot.png | Bin 35497 -> 63551 bytes .../mqc_deeptools_plotFingerprint_plot.png | Bin 114836 -> 78044 bytes .../images/mqc_deeptools_plotProfile_plot.png | Bin 115914 -> 74083 bytes docs/images/mqc_deseq2_pca_plot.png | Bin 61615 -> 21263 bytes .../mqc_deseq2_sample_similarity_plot.png | Bin 76247 -> 25642 bytes .../mqc_featureCounts_assignment_plot.png | Bin 76240 -> 26436 bytes docs/images/mqc_frip_score_plot.png | Bin 74548 -> 26374 bytes docs/images/mqc_macs2_peak_count_plot.png | Bin 72526 -> 26994 bytes docs/images/mqc_picard_deduplication_plot.png | Bin 76669 -> 27788 bytes docs/images/mqc_picard_insert_size_plot.png | Bin 116371 -> 59566 bytes docs/images/mqc_preseq_plot.png | Bin 108852 -> 53270 bytes 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| 4 +++- 1 file changed, 3 insertions(+), 1 deletion(-) diff --git a/docs/output.md b/docs/output.md index 241f7abe..9c6d8a6d 100755 --- a/docs/output.md +++ b/docs/output.md @@ -223,7 +223,9 @@ This pipeline uses a standardised DESeq2 analysis script to get an idea of the r ![MultiQC - DESeq2 PCA plot](images/mqc_deseq2_pca_plot.png) -![MultiQC - DESeq2 sample similarity plot](images/mqc_deseq2_sample_similarity_plot.png) +

+ MultiQC - DESeq2 sample similarity plot +

By default, all possible pairwise comparisons across the groups are performed (as defined in [`design.csv`](usage.md#--design)). The DESeq2 results are generated by the pipeline in various ways. You can load up the results across all of the comparisons in a single spreadsheet, or individual folders will also be created that contain the results specific to a particular comparison. For the latter, additional files will also be generated where the intervals have been pre-filtered based on a couple of standard FDR thresholds. Please see [DESeq2 output](http://bioconductor.org/packages/release/bioc/vignettes/DESeq2/inst/doc/DESeq2.html#differential-expression-analysis) for a description of the columns generated by DESeq2. From 07b09a8e61f71f3d11068dd5c7275516cca329ad Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 30 Jun 2020 18:48:03 +0100 Subject: [PATCH 212/226] Resize image --- docs/output.md | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/docs/output.md b/docs/output.md index 9c6d8a6d..ce2063bc 100755 --- a/docs/output.md +++ b/docs/output.md @@ -223,8 +223,8 @@ This pipeline uses a standardised DESeq2 analysis script to get an idea of the r ![MultiQC - DESeq2 PCA plot](images/mqc_deseq2_pca_plot.png) -

- MultiQC - DESeq2 sample similarity plot +

+ MultiQC - DESeq2 sample similarity plot

By default, all possible pairwise comparisons across the groups are performed (as defined in [`design.csv`](usage.md#--design)). The DESeq2 results are generated by the pipeline in various ways. You can load up the results across all of the comparisons in a single spreadsheet, or individual folders will also be created that contain the results specific to a particular comparison. For the latter, additional files will also be generated where the intervals have been pre-filtered based on a couple of standard FDR thresholds. Please see [DESeq2 output](http://bioconductor.org/packages/release/bioc/vignettes/DESeq2/inst/doc/DESeq2.html#differential-expression-analysis) for a description of the columns generated by DESeq2. From c56c7927368f49399159fc406922c84f2dbf1f5c Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 30 Jun 2020 18:48:34 +0100 Subject: [PATCH 213/226] Resize image --- docs/output.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/docs/output.md b/docs/output.md index ce2063bc..47edf763 100755 --- a/docs/output.md +++ b/docs/output.md @@ -224,7 +224,7 @@ This pipeline uses a standardised DESeq2 analysis script to get an idea of the r ![MultiQC - DESeq2 PCA plot](images/mqc_deseq2_pca_plot.png)

- MultiQC - DESeq2 sample similarity plot + MultiQC - DESeq2 sample similarity plot

By default, all possible pairwise comparisons across the groups are performed (as defined in [`design.csv`](usage.md#--design)). The DESeq2 results are generated by the pipeline in various ways. You can load up the results across all of the comparisons in a single spreadsheet, or individual folders will also be created that contain the results specific to a particular comparison. For the latter, additional files will also be generated where the intervals have been pre-filtered based on a couple of standard FDR thresholds. Please see [DESeq2 output](http://bioconductor.org/packages/release/bioc/vignettes/DESeq2/inst/doc/DESeq2.html#differential-expression-analysis) for a description of the columns generated by DESeq2. From 9f1a5fec22e25798e546e6faf27ecc7408941d5b Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 30 Jun 2020 18:49:09 +0100 Subject: [PATCH 214/226] Resize image --- docs/output.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/docs/output.md b/docs/output.md index 47edf763..7e89a9df 100755 --- a/docs/output.md +++ b/docs/output.md @@ -224,7 +224,7 @@ This pipeline uses a standardised DESeq2 analysis script to get an idea of the r ![MultiQC - DESeq2 PCA plot](images/mqc_deseq2_pca_plot.png)

- MultiQC - DESeq2 sample similarity plot + MultiQC - DESeq2 sample similarity plot

By default, all possible pairwise comparisons across the groups are performed (as defined in [`design.csv`](usage.md#--design)). The DESeq2 results are generated by the pipeline in various ways. You can load up the results across all of the comparisons in a single spreadsheet, or individual folders will also be created that contain the results specific to a particular comparison. For the latter, additional files will also be generated where the intervals have been pre-filtered based on a couple of standard FDR thresholds. Please see [DESeq2 output](http://bioconductor.org/packages/release/bioc/vignettes/DESeq2/inst/doc/DESeq2.html#differential-expression-analysis) for a description of the columns generated by DESeq2. From 5e77dd90548d1ad369c0b9d8f5ade522daa8a5b6 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 1 Jul 2020 01:14:02 +0100 Subject: [PATCH 215/226] Update MultiQC header files --- assets/multiqc/mlib_deseq2_clustering_header.txt | 4 ++-- assets/multiqc/mlib_deseq2_pca_header.txt | 4 ++-- assets/multiqc/mlib_frip_score_header.txt | 4 ++-- assets/multiqc/mlib_peak_annotation_header.txt | 4 ++-- assets/multiqc/mlib_peak_count_header.txt | 4 ++-- assets/multiqc/mrep_deseq2_clustering_header.txt | 4 ++-- assets/multiqc/mrep_deseq2_pca_header.txt | 4 ++-- assets/multiqc/mrep_frip_score_header.txt | 4 ++-- assets/multiqc/mrep_peak_annotation_header.txt | 4 ++-- assets/multiqc/mrep_peak_count_header.txt | 4 ++-- 10 files changed, 20 insertions(+), 20 deletions(-) diff --git a/assets/multiqc/mlib_deseq2_clustering_header.txt b/assets/multiqc/mlib_deseq2_clustering_header.txt index 60237214..be3b8085 100644 --- a/assets/multiqc/mlib_deseq2_clustering_header.txt +++ b/assets/multiqc/mlib_deseq2_clustering_header.txt @@ -1,11 +1,11 @@ #id: 'mlib_deseq2_clustering' -#section_name: 'DESeq2: Sample similarity (merged library)' +#section_name: 'MERGED LIB: DESeq2 sample similarity' #description: "is generated from clustering by Euclidean distances between # DESeq2 # rlog values for each sample # in the featurecounts_deseq2.r script." #plot_type: 'heatmap' -#anchor: 'nfcore_atacseq-mlib_deseq2_clustering' +#anchor: 'mlib_deseq2_clustering' #pconfig: # title: 'DESeq2: Heatmap of the sample-to-sample distances' # xlab: True diff --git a/assets/multiqc/mlib_deseq2_pca_header.txt b/assets/multiqc/mlib_deseq2_pca_header.txt index 434c305d..ba6be8e2 100644 --- a/assets/multiqc/mlib_deseq2_pca_header.txt +++ b/assets/multiqc/mlib_deseq2_pca_header.txt @@ -1,10 +1,10 @@ #id: 'mlib_deseq2_pca' -#section_name: 'DESeq2: PCA plot (merged library)' +#section_name: 'MERGED LIB: DESeq2 PCA plot' #description: "PCA plot between samples in the experiment. # These values are calculated using DESeq2 # in the featurecounts_deseq2.r script." #plot_type: 'scatter' -#anchor: 'nfcore_atacseq-mlib_deseq2_pca' +#anchor: 'mlib_deseq2_pca' #pconfig: # title: 'DESeq2: Principal component plot' # xlab: PC1 diff --git a/assets/multiqc/mlib_frip_score_header.txt b/assets/multiqc/mlib_frip_score_header.txt index 36fb7e8e..d0edf803 100644 --- a/assets/multiqc/mlib_frip_score_header.txt +++ b/assets/multiqc/mlib_frip_score_header.txt @@ -1,10 +1,10 @@ #id: 'mlib_frip_score' -#section_name: 'MACS2: Peak FRiP score (merged library)' +#section_name: 'MERGED LIB: MACS2 peak FRiP score' #description: "is generated by calculating the fraction of all mapped reads that fall # into the MACS2 called peak regions. A read must overlap a peak by at least 20% to be counted. # See FRiP score." #plot_type: 'bargraph' -#anchor: 'nfcore_atacseq-mlib_frip_score' +#anchor: 'mlib_frip_score' #pconfig: # title: 'FRiP score' # ylab: 'FRiP score' diff --git a/assets/multiqc/mlib_peak_annotation_header.txt b/assets/multiqc/mlib_peak_annotation_header.txt index 3abac2f0..6f668c31 100644 --- a/assets/multiqc/mlib_peak_annotation_header.txt +++ b/assets/multiqc/mlib_peak_annotation_header.txt @@ -1,9 +1,9 @@ #id: 'mlib_peak_annotation' -#section_name: 'HOMER: Peak annotation (merged library)' +#section_name: 'MERGED LIB: HOMER peak annotation' #description: "is generated by calculating the proportion of peaks assigned to genomic features by # HOMER annotatePeaks.pl." #plot_type: 'bargraph' -#anchor: 'nfcore_atacseq-mlib_peak_annotation' +#anchor: 'mlib_peak_annotation' #pconfig: # title: 'Peak to feature proportion' # ylab: 'Peak count' diff --git a/assets/multiqc/mlib_peak_count_header.txt b/assets/multiqc/mlib_peak_count_header.txt index 31f03b72..9fd8c830 100644 --- a/assets/multiqc/mlib_peak_count_header.txt +++ b/assets/multiqc/mlib_peak_count_header.txt @@ -1,9 +1,9 @@ #id: 'mlib_peak_count' -#section_name: 'MACS2: Peak count (merged library)' +#section_name: 'MERGED LIB: MACS2 peak count' #description: "is calculated from total number of peaks called by # MACS2" #plot_type: 'bargraph' -#anchor: 'nfcore_atacseq-mlib_peak_count' +#anchor: 'mlib_peak_count' #pconfig: # title: 'Total peak count' # ylab: 'Peak count' diff --git a/assets/multiqc/mrep_deseq2_clustering_header.txt b/assets/multiqc/mrep_deseq2_clustering_header.txt index b1fdafe9..d36e8eab 100644 --- a/assets/multiqc/mrep_deseq2_clustering_header.txt +++ b/assets/multiqc/mrep_deseq2_clustering_header.txt @@ -1,11 +1,11 @@ #id: 'mrep_deseq2_clustering' -#section_name: 'DESeq2: Sample similarity (merged replicate)' +#section_name: 'MERGED REP: DESeq2 sample similarity' #description: "is generated from clustering by Euclidean distances between # DESeq2 # rlog values for each sample # in the featurecounts_deseq2.r script." #plot_type: 'heatmap' -#anchor: 'nfcore_atacseq-mrep_deseq2_clustering' +#anchor: 'mrep_deseq2_clustering' #pconfig: # title: 'DESeq2: Heatmap of the sample-to-sample distances' # xlab: True diff --git a/assets/multiqc/mrep_deseq2_pca_header.txt b/assets/multiqc/mrep_deseq2_pca_header.txt index 1ba35a93..2f58a377 100644 --- a/assets/multiqc/mrep_deseq2_pca_header.txt +++ b/assets/multiqc/mrep_deseq2_pca_header.txt @@ -1,10 +1,10 @@ #id: 'mrep_deseq2_pca' -#section_name: 'DESeq2: PCA plot (merged replicate)' +#section_name: 'MERGED REP: DESeq2 PCA plot' #description: "PCA plot between samples in the experiment. # These values are calculated using DESeq2 # in the featurecounts_deseq2.r script." #plot_type: 'scatter' -#anchor: 'nfcore_atacseq-mrep_deseq2_pca' +#anchor: 'mrep_deseq2_pca' #pconfig: # title: 'DESeq2: Principal component plot' # xlab: PC1 diff --git a/assets/multiqc/mrep_frip_score_header.txt b/assets/multiqc/mrep_frip_score_header.txt index 92265b06..8a2c5ad1 100644 --- a/assets/multiqc/mrep_frip_score_header.txt +++ b/assets/multiqc/mrep_frip_score_header.txt @@ -1,10 +1,10 @@ #id: 'mrep_frip_score' -#section_name: 'MACS2: Peak FRiP score (merged replicate)' +#section_name: 'MERGED REP: MACS2 peak FRiP score' #description: "is generated by calculating the fraction of all mapped reads that fall # into the MACS2 called peak regions. A read must overlap a peak by at least 20% to be counted. # See FRiP score." #plot_type: 'bargraph' -#anchor: 'nfcore_atacseq-mrep_frip_score' +#anchor: 'mrep_frip_score' #pconfig: # title: 'FRiP score' # ylab: 'FRiP score' diff --git a/assets/multiqc/mrep_peak_annotation_header.txt b/assets/multiqc/mrep_peak_annotation_header.txt index 4e605e68..d9d0eb03 100644 --- a/assets/multiqc/mrep_peak_annotation_header.txt +++ b/assets/multiqc/mrep_peak_annotation_header.txt @@ -1,9 +1,9 @@ #id: 'mrep_peak_annotation' -#section_name: 'HOMER: Peak annotation (merged replicate)' +#section_name: 'MERGED REP: HOMER peak annotation' #description: "is generated by calculating the proportion of peaks assigned to genomic features by # HOMER annotatePeaks.pl." #plot_type: 'bargraph' -#anchor: 'nfcore_atacseq-mrep_peak_annotation' +#anchor: 'mrep_peak_annotation' #pconfig: # title: 'Peak to feature proportion' # ylab: 'Peak count' diff --git a/assets/multiqc/mrep_peak_count_header.txt b/assets/multiqc/mrep_peak_count_header.txt index 2d69a0aa..66796ffe 100644 --- a/assets/multiqc/mrep_peak_count_header.txt +++ b/assets/multiqc/mrep_peak_count_header.txt @@ -1,9 +1,9 @@ #id: 'mrep_peak_count' -#section_name: 'MACS2: Peak count (merged replicate)' +#section_name: 'MERGED REP: MACS2 Peak count' #description: "is calculated from total number of peaks called by # MACS2" #plot_type: 'bargraph' -#anchor: 'nfcore_atacseq-mrep_peak_count' +#anchor: 'mrep_peak_count' #pconfig: # title: 'Total peak count' # ylab: 'Peak count' From 7109c10d7a274a7c52fe18c4e3c100741056b7f9 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 1 Jul 2020 01:14:13 +0100 Subject: [PATCH 216/226] Fix Picard version --- bin/scrape_software_versions.py | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/bin/scrape_software_versions.py b/bin/scrape_software_versions.py index 022a8758..585d1805 100755 --- a/bin/scrape_software_versions.py +++ b/bin/scrape_software_versions.py @@ -13,7 +13,7 @@ 'BEDTools': ['v_bedtools.txt', r"bedtools v(\S+)"], 'BamTools': ['v_bamtools.txt', r"bamtools (\S+)"], 'deepTools': ['v_deeptools.txt', r"plotFingerprint (\S+)"], - 'Picard': ['v_picard.txt', r"([\d\.]+)-SNAPSHOT"], + 'Picard': ['v_picard.txt', r"\n(\S+)"], 'R': ['v_R.txt', r"R version (\S+)"], 'Pysam': ['v_pysam.txt', r"(\S+)"], 'MACS2': ['v_macs2.txt', r"macs2 (\S+)"], From 67e62fcb4331d982db2d08b44b080d17b46548a7 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 1 Jul 2020 01:14:32 +0100 Subject: [PATCH 217/226] TLC for MultiQC --- assets/multiqc_config.yaml | 123 ++++++++++++++++++++----------------- 1 file changed, 68 insertions(+), 55 deletions(-) diff --git a/assets/multiqc_config.yaml b/assets/multiqc_config.yaml index 5d07c929..9416fbf1 100755 --- a/assets/multiqc_config.yaml +++ b/assets/multiqc_config.yaml @@ -20,100 +20,98 @@ exclude_modules: module_order: - fastqc: - name: 'FastQC (library; raw)' - info: 'This section of the report shows FastQC results before adapter trimming.' + name: 'LIB: FastQC (raw)' + info: 'This section of the report shows FastQC results before adapter trimming for individual libraries.' path_filters: - - '*_fastqc.zip' - path_filters_exclude: - - '*val*_fastqc.zip' - - '*trimmed_fastqc.zip' + - './fastqc/*.zip' - cutadapt: - name: 'cutadapt (library; trimmed)' - info: 'This section of the report shows the length of trimmed reads by cutadapt.' - path_filters: - - '*_trimming_report.txt' + name: 'LIB: Cutadapt (trimmed)' + info: 'This section of the report shows the length of trimmed reads by Cutadapt for individual libraries.' - fastqc: - name: 'FastQC (library; trimmed)' - info: 'This section of the report shows FastQC results after adapter trimming.' + name: 'LIB: FastQC (trimmed)' + info: 'This section of the report shows FastQC results after adapter trimming for individual libraries.' path_filters: - - '*val*_fastqc.zip' - - '*trimmed_fastqc.zip' + - './trimgalore/fastqc/*.zip' - samtools: - name: 'SAMTools (library)' + name: 'LIB: SAMTools' info: 'This section of the report shows SAMTools results for individual libraries.' path_filters: - - '*.Lb.sorted.bam*' + - './alignment/library/*' - samtools: - name: 'SAMTools (merged library; unfiltered)' + name: 'MERGED LIB: SAMTools (unfiltered)' info: 'This section of the report shows SAMTools results after merging libraries and before filtering.' path_filters: - - '*mLb.mkD.sorted.bam*' + - './alignment/mergedLibrary/*.mLb.mkD.sorted.bam*' - preseq: - name: 'Preseq (merged library; unfiltered)' + name: 'MERGED LIB: Preseq (unfiltered)' info: 'This section of the report shows Preseq results after merging libraries and before filtering.' - path_filters: - - '*mLb*' - samtools: - name: 'SAMTools (merged library; filtered)' + name: 'MERGED LIB: SAMTools (filtered)' info: 'This section of the report shows SAMTools results after merging libraries and after filtering.' path_filters: - - '*mLb.clN.sorted.bam*' + - './alignment/mergedLibrary/*.mLb.clN.sorted.bam*' - picard: - name: 'Picard (merged library; filtered)' + name: 'MERGED LIB: Picard' info: 'This section of the report shows picard results after merging libraries and after filtering.' path_filters: - - '*mLb*' + - './alignment/mergedLibrary/picard_metrics/*' - deeptools: - name: 'deepTools' + name: 'MERGED LIB: deepTools' + anchor: 'mlib_deeptools' info: 'This section of the report shows QC plots generated by deepTools.' + - featureCounts: + name: 'MERGED LIB: featureCounts' + anchor: 'mlib_featurecounts' + info: 'This section of the report shows featureCounts results for the number of reads assigned to merged library consensus peaks.' path_filters: - - '*.plot*' + - './macs/mergedLibrary/consensus/*.summary' - samtools: - name: 'SAMTools (merged replicate; filtered)' - info: 'This section of the report shows SAMTools results after merging replicates and before filtering.' + name: 'MERGED REP: SAMTools' + info: 'This section of the report shows SAMTools results after merging replicates and filtering.' path_filters: - - '*mRp.clN*' + - './alignment/mergedReplicate/*' - picard: - name: 'Picard (merged replicate; filtered)' + name: 'MERGED REP: Picard' + anchor: 'mrep_picard' info: 'This section of the report shows picard results after merging libraries and before filtering.' path_filters: - - '*mRp.clN*' + - './alignment/mergedReplicate/*' - featureCounts: - name: 'featureCounts (merged library)' - info: 'This section of the report shows featureCounts results for the number of reads assigned to merged library consensus peaks.' - path_filters: - - '*mLb.clN.featureCounts*' - - featureCounts: - name: 'featureCounts (merged replicate)' + name: 'MERGED REP: featureCounts' + anchor: 'mrep_featurecounts' info: 'This section of the report shows featureCounts results for the number of reads assigned to merged replicate consensus peaks.' path_filters: - - '*mRp.clN.featureCounts*' + - './macs/mergedReplicate/consensus/*.summary' report_section_order: mlib_peak_count: - order: -1000 - mrep_peak_count: - order: -1100 + before: mlib_deeptools mlib_frip_score: - order: -1200 - mrep_frip_score: - order: -1300 + before: mlib_peak_count mlib_peak_annotation: - order: -1400 - mrep_peak_annotation: - order: -1500 + before: mlib_frip_score + mlib_featurecounts: + before: mlib_peak_annotation mlib_deseq2_pca: - order: -1600 - mrep_deseq2_pca: - order: -1700 + before: mlib_featurecounts mlib_deseq2_clustering: - order: -1800 + before: mlib_deseq2_pca + mrep_peak_count: + before: mrep_picard + mrep_frip_score: + before: mrep_peak_count + mrep_peak_annotation: + before: mrep_frip_score + mrep_featurecounts: + before: mrep_peak_annotation + mrep_deseq2_pca: + before: mrep_featurecounts mrep_deseq2_clustering: - order: -1900 + before: mrep_deseq2_pca software_versions: - order: -2000 + order: -1001 nf-core-atacseq-summary: - order: -2100 + order: -1002 custom_plot_config: picard_insert_size: @@ -135,3 +133,18 @@ extra_fn_clean_exts: - 'mLb' - 'mRp' - '_peaks' + +# # Customise the module search patterns to speed up execution time +# # - Skip module sub-tools that we are not interested in +# # - Replace file-content searching with filename pattern searching +# # - Don't add anything that is the same as the MultiQC default +# # See https://multiqc.info/docs/#optimise-file-search-patterns for details +sp: + cutadapt: + fn: '*trimming_report.txt' + preseq: + fn: '*.ccurve.txt' + deeptools/plotFingerprintOutRawCounts: + fn: '*plotFingerprint*' + deeptools/plotProfile: + fn: '*plotProfile*' From 4ca0173d6dd07aaa4c18cf09272d217fdb70b2a0 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 1 Jul 2020 13:09:57 +0100 Subject: [PATCH 218/226] Fix #109 --- main.nf | 10 ++++++++++ 1 file changed, 10 insertions(+) diff --git a/main.nf b/main.nf index 291de879..fede726e 100644 --- a/main.nf +++ b/main.nf @@ -409,7 +409,17 @@ if (!params.bwa_index) { /* * PREPROCESSING: Generate gene BED file */ +// If --gtf is supplied along with --genome +// Make gene bed from supplied --gtf instead of using iGenomes one automatically +def MAKE_BED = false if (!params.gene_bed) { + MAKE_BED = true +} else if (params.genome && params.gtf) { + if (params.genomes[ params.genome ].gtf != params.gtf) { + MAKE_BED = true + } +} +if (MAKE_BED) { process MAKE_GENE_BED { tag "$gtf" label 'process_low' From 6260e65020d7dc2faf2d9cffec4fc737f4a879b4 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 1 Jul 2020 13:19:09 +0100 Subject: [PATCH 219/226] Update CHANGELOG --- CHANGELOG.md | 2 ++ 1 file changed, 2 insertions(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index 21f5a30d..08123cc3 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -14,7 +14,9 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * [#79](https://github.com/nf-core/atacseq/issues/79) - Parallelize DESeq2 * [#80](https://github.com/nf-core/atacseq/pull/80) - Added social preview image * [#97](https://github.com/nf-core/atacseq/issues/97) - PBC1, PBC2 from pipeline? +* [#109](https://github.com/nf-core/atacseq/issues/109) - Specify custom gtf but gene bed is not generated from that gtf? * [nf-core/chipseq#153](https://github.com/nf-core/chipseq/issues/153) - Add plotHeatmap +* Regenerated screenshots and added collapsible sections for output files in `docs/output.md` * Update template to tools `1.9` * Replace `set` with `tuple` and `file()` with `path()` in all processes * Capitalise process names From 1063986f0163163e8b9b8bccc2b2adfcd255088d Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 1 Jul 2020 13:42:25 +0100 Subject: [PATCH 220/226] Fix #107 --- CHANGELOG.md | 3 +++ docs/usage.md | 10 ++++++++++ main.nf | 8 ++++++++ nextflow.config | 2 ++ 4 files changed, 23 insertions(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index 08123cc3..f6f87c76 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -14,6 +14,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * [#79](https://github.com/nf-core/atacseq/issues/79) - Parallelize DESeq2 * [#80](https://github.com/nf-core/atacseq/pull/80) - Added social preview image * [#97](https://github.com/nf-core/atacseq/issues/97) - PBC1, PBC2 from pipeline? +* [#107](https://github.com/nf-core/atacseq/issues/107) - Add options to change MACS2 parameters * [#109](https://github.com/nf-core/atacseq/issues/109) - Specify custom gtf but gene bed is not generated from that gtf? * [nf-core/chipseq#153](https://github.com/nf-core/chipseq/issues/153) - Add plotHeatmap * Regenerated screenshots and added collapsible sections for output files in `docs/output.md` @@ -21,6 +22,8 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * Replace `set` with `tuple` and `file()` with `path()` in all processes * Capitalise process names * Parameters: + * `--macs_fdr` to provide FDR threshold for MACS2 peak calling + * `--macs_pvalue` to provide p-value threshold for MACS2 peak calling * `--skip_peak_qc` to skip MACS2 peak QC plot generation * `--skip_peak_annotation` to skip annotation of MACS2 and consensus peaks with HOMER * `--skip_consensus_peaks` to skip consensus peak generation diff --git a/docs/usage.md b/docs/usage.md index 41eff287..ec4dbf8e 100755 --- a/docs/usage.md +++ b/docs/usage.md @@ -39,6 +39,8 @@ * [Peaks](#peaks) * [`--narrow_peak`](#--narrow_peak) * [`--broad_cutoff`](#--broad_cutoff) + * [`--macs_fdr`](#--macs_fdr) + * [`--macs_pvalue`](#--macs_pvalue) * [`--min_reps_consensus`](#--min_reps_consensus) * [`--save_macs_pileup`](#--save_macs_pileup) * [`--skip_peak_qc`](#--skip_peak_qc) @@ -388,6 +390,14 @@ MACS2 is run by default with the [`--broad`](https://github.com/taoliu/MACS#--br Specifies broad cut-off value for MACS2. Only used when `--narrow_peak` isnt specified (Default: `0.1`). +### `--macs_fdr` + +Minimum FDR (q-value) cutoff for peak detection, `--macs_fdr` and `--macs_pvalue` are mutually exclusive (Default: false). + +### `--macs_pvalue` + +p-value cutoff for peak detection, `--macs_fdr` and `--macs_pvalue` are mutually exclusive (Default: false). If `--macs_pvalue` cutoff is set, q-value will not be calculated and reported as -1 in the final .xls file. + ### `--min_reps_consensus` Number of biological replicates required from a given condition for a peak to contribute to a consensus peak . If you are confident you have good reproducibility amongst your replicates then you can increase the value of this parameter to create a "reproducible" set of consensus of peaks. For example, a value of 2 will mean peaks that have been called in at least 2 replicates will contribute to the consensus set of peaks, and as such peaks that are unique to a given replicate will be discarded. diff --git a/main.nf b/main.nf index fede726e..0f2e4f34 100644 --- a/main.nf +++ b/main.nf @@ -60,6 +60,8 @@ def helpMessage() { Peaks --narrow_peak [bool] Run MACS2 in narrowPeak mode (Default: false) --broad_cutoff [float] Specifies broad cutoff value for MACS2. Only used when --narrow_peak isnt specified (Default: 0.1) + --macs_fdr [float] Minimum FDR (q-value) cutoff for peak detection, --macs_fdr and --macs_pvalue are mutually exclusive (Default: false) + --macs_pvalue [float] p-value cutoff for peak detection, --macs_fdr and --macs_pvalue are mutually exclusive (Default: false) --min_reps_consensus [int] Number of biological replicates required from a given condition for a peak to contribute to a consensus peak (Default: 1) --save_macs_pileup [bool] Instruct MACS2 to create bedGraph files normalised to signal per million reads (Default: false) --skip_peak_qc [bool] Skip MACS2 peak QC plot generation (Default: false) @@ -241,6 +243,8 @@ summary['MACS2 Genome Size'] = params.macs_gsize ?: 'Not supplied' summary['Min Consensus Reps'] = params.min_reps_consensus if (params.macs_gsize) summary['MACS2 Narrow Peaks'] = params.narrow_peak ? 'Yes' : 'No' if (!params.narrow_peak) summary['MACS2 Broad Cutoff'] = params.broad_cutoff +if (params.macs_fdr) summary['MACS2 FDR'] = params.macs_fdr +if (params.macs_pvalue) summary['MACS2 P-value'] = params.macs_pvalue if (params.skip_trimming) { summary['Trimming Step'] = 'Skipped' } else { @@ -1131,6 +1135,8 @@ process MERGED_LIB_MACS2 { broad = params.narrow_peak ? '' : "--broad --broad-cutoff ${params.broad_cutoff}" format = params.single_end ? 'BAM' : 'BAMPE' pileup = params.save_macs_pileup ? '-B --SPMR' : '' + fdr = params.macs_fdr ? "--qvalue ${params.macs_fdr}" : '' + pvalue = params.macs_pvalue ? "--pvalue ${params.macs_pvalue}" : '' """ macs2 callpeak \\ -t ${bam[0]} \\ @@ -1139,6 +1145,8 @@ process MERGED_LIB_MACS2 { -g $params.macs_gsize \\ -n $prefix \\ $pileup \\ + $fdr \\ + $pvalue \\ --keep-dup all \\ --nomodel diff --git a/nextflow.config b/nextflow.config index aafd8841..4a2e03ce 100644 --- a/nextflow.config +++ b/nextflow.config @@ -39,6 +39,8 @@ params { // Options: Peaks narrow_peak = false broad_cutoff = 0.1 + macs_fdr = false + macs_pvalue = false min_reps_consensus = 1 save_macs_pileup = false skip_peak_qc = false From 335251b140ee9fa7a5fa347d08a34254eb4ee534 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 1 Jul 2020 14:30:26 +0100 Subject: [PATCH 221/226] Fix chipseq#159 --- CHANGELOG.md | 2 ++ docs/usage.md | 39 ++++++++++++++++++++++----------------- main.nf | 4 ++++ nextflow.config | 1 + 4 files changed, 29 insertions(+), 17 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index f6f87c76..36a10e05 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -17,11 +17,13 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * [#107](https://github.com/nf-core/atacseq/issues/107) - Add options to change MACS2 parameters * [#109](https://github.com/nf-core/atacseq/issues/109) - Specify custom gtf but gene bed is not generated from that gtf? * [nf-core/chipseq#153](https://github.com/nf-core/chipseq/issues/153) - Add plotHeatmap +* [nf-core/chipseq#159](https://github.com/nf-core/chipseq/issues/159) - expose bwa mem -T parameter * Regenerated screenshots and added collapsible sections for output files in `docs/output.md` * Update template to tools `1.9` * Replace `set` with `tuple` and `file()` with `path()` in all processes * Capitalise process names * Parameters: + * `--bwa_min_score` to set minimum alignment score for BWA MEM * `--macs_fdr` to provide FDR threshold for MACS2 peak calling * `--macs_pvalue` to provide p-value threshold for MACS2 peak calling * `--skip_peak_qc` to skip MACS2 peak QC plot generation diff --git a/docs/usage.md b/docs/usage.md index ec4dbf8e..f772ef50 100755 --- a/docs/usage.md +++ b/docs/usage.md @@ -31,6 +31,7 @@ * [`--skip_trimming`](#--skip_trimming) * [`--save_trimmed`](#--save_trimmed) * [Alignments](#alignments) + * [`--bwa_min_score`](#--bwa_min_score) * [`--keep_mito`](#--keep_mito) * [`--keep_dups`](#--keep_dups) * [`--keep_multi_map`](#--keep_multi_map) @@ -328,11 +329,11 @@ Name of mitochondrial chomosome in reference assembly. Reads aligning to this co ### `--save_reference` -If the BWA index is generated by the pipeline use this parameter to save it to your results folder. These can then be used for future pipeline runs, reducing processing times. +If the BWA index is generated by the pipeline use this parameter to save it to your results folder. These can then be used for future pipeline runs, reducing processing times (Default: false). ### `--igenomes_ignore` -Do not load `igenomes.config` when running the pipeline. You may choose this option if you observe clashes between custom parameters and those supplied in `igenomes.config`. +Do not load `igenomes.config` when running the pipeline. You may choose this option if you observe clashes between custom parameters and those supplied in `igenomes.config` (Default: false). ## Adapter trimming @@ -352,39 +353,43 @@ You can specify custom trimming parameters as follows: ### `--skip_trimming` -Skip the adapter trimming step. Use this if your input FastQ files have already been trimmed outside of the workflow or if you're very confident that there is no adapter contamination in your data. +Skip the adapter trimming step. Use this if your input FastQ files have already been trimmed outside of the workflow or if you're very confident that there is no adapter contamination in your data (Default: false). ### `--save_trimmed` -By default, trimmed FastQ files will not be saved to the results directory. Specify this flag (or set to true in your config file) to copy these files to the results directory when complete. +By default, trimmed FastQ files will not be saved to the results directory. Specify this flag (or set to true in your config file) to copy these files to the results directory when complete (Default: false). ## Alignments +### `--bwa_min_score` + +Don’t output BWA MEM alignments with score lower than this parameter (Default: false). + ### `--keep_mito` -Reads mapping to mitochondrial contig are not filtered from alignments. +Reads mapping to mitochondrial contig are not filtered from alignments (Default: false). ### `--keep_dups` -Duplicate reads are not filtered from alignments. +Duplicate reads are not filtered from alignments (Default: false). ### `--keep_multi_map` -Reads mapping to multiple locations in the genome are not filtered from alignments. +Reads mapping to multiple locations in the genome are not filtered from alignments (Default: false). ### `--skip_merge_replicates` -An additional series of steps are performed by the pipeline by merging the replicates from the same experimental group. This is primarily to increase the sequencing depth in order to perform downstream analyses such as footprinting. Specifying this parameter means that these steps will not be performed. +An additional series of steps are performed by the pipeline by merging the replicates from the same experimental group. This is primarily to increase the sequencing depth in order to perform downstream analyses such as footprinting. Specifying this parameter means that these steps will not be performed (Default: false). ### `--save_align_intermeds` -By default, intermediate BAM files will not be saved. The final BAM files created after the appropriate filtering step are always saved to limit storage usage. Set to true to also save other intermediate BAM files. +By default, intermediate BAM files will not be saved. The final BAM files created after the appropriate filtering step are always saved to limit storage usage. Set to true to also save other intermediate BAM files (Default: false). ## Peaks ### `--narrow_peak` -MACS2 is run by default with the [`--broad`](https://github.com/taoliu/MACS#--broad) flag. Specify this flag to call peaks in narrowPeak mode. +MACS2 is run by default with the [`--broad`](https://github.com/taoliu/MACS#--broad) flag. Specify this flag to call peaks in narrowPeak mode (Default: false). ### `--broad_cutoff` @@ -400,7 +405,7 @@ p-value cutoff for peak detection, `--macs_fdr` and `--macs_pvalue` are mutually ### `--min_reps_consensus` -Number of biological replicates required from a given condition for a peak to contribute to a consensus peak . If you are confident you have good reproducibility amongst your replicates then you can increase the value of this parameter to create a "reproducible" set of consensus of peaks. For example, a value of 2 will mean peaks that have been called in at least 2 replicates will contribute to the consensus set of peaks, and as such peaks that are unique to a given replicate will be discarded. +Number of biological replicates required from a given condition for a peak to contribute to a consensus peak . If you are confident you have good reproducibility amongst your replicates then you can increase the value of this parameter to create a "reproducible" set of consensus of peaks. For example, a value of 2 will mean peaks that have been called in at least 2 replicates will contribute to the consensus set of peaks, and as such peaks that are unique to a given replicate will be discarded (Default: 1). ```bash -- min_reps_consensus 1 @@ -408,29 +413,29 @@ Number of biological replicates required from a given condition for a peak to co ### `--save_macs_pileup` -Instruct MACS2 to create bedGraph files using the `-B --SPMR` parameters. +Instruct MACS2 to create bedGraph files using the `-B --SPMR` parameters (Default: false). ### `--skip_peak_qc` -Skip MACS2 peak QC plot generation. +Skip MACS2 peak QC plot generation (Default: false). ### `--skip_peak_annotation` -Skip annotation of MACS2 and consensus peaks with HOMER. +Skip annotation of MACS2 and consensus peaks with HOMER (Default: false). ### `--skip_consensus_peaks` -Skip consensus peak generation, annotation and counting. +Skip consensus peak generation, annotation and counting (Default: false). ## Differential analysis ### `--deseq2_vst` -Use `vst` transformation instead of `rlog` with DESeq2. See [DESeq2 docs](http://bioconductor.org/packages/devel/bioc/vignettes/DESeq2/inst/doc/DESeq2.html#data-transformations-and-visualization). +Use `vst` transformation instead of `rlog` with DESeq2. See [DESeq2 docs](http://bioconductor.org/packages/devel/bioc/vignettes/DESeq2/inst/doc/DESeq2.html#data-transformations-and-visualization) (Default: false). ### `--skip_diff_analysis` -Skip differential accessibility analysis with DESeq2. +Skip differential accessibility analysis with DESeq2 (Default: false). ## Skipping QC steps diff --git a/main.nf b/main.nf index 0f2e4f34..44e4b9b0 100644 --- a/main.nf +++ b/main.nf @@ -51,6 +51,7 @@ def helpMessage() { --save_trimmed [bool] Save the trimmed FastQ files in the results directory (Default: false) Alignments + --bwa_min_score [int] Don’t output BWA MEM alignments with score lower than this parameter (Default: false) --keep_mito [bool] Reads mapping to mitochondrial contig are not filtered from alignments (Default: false) --keep_dups [bool] Duplicate reads are not filtered from alignments (Default: false) --keep_multi_map [bool] Reads mapping to multiple locations are not filtered from alignments (Default: false) @@ -239,6 +240,7 @@ if (params.tss_bed) summary['TSS BED File'] = params.tss_bed if (params.bwa_index) summary['BWA Index'] = params.bwa_index if (params.blacklist) summary['Blacklist BED'] = params.blacklist if (params.mito_name) summary['Mitochondrial Contig'] = params.mito_name +if (params.bwa_min_score) summary['BWA Min Score'] = params.bwa_min_score summary['MACS2 Genome Size'] = params.macs_gsize ?: 'Not supplied' summary['Min Consensus Reps'] = params.min_reps_consensus if (params.macs_gsize) summary['MACS2 Narrow Peaks'] = params.narrow_peak ? 'Yes' : 'No' @@ -637,11 +639,13 @@ process BWA_MEM { if (params.seq_center) { rg = "\'@RG\\tID:${name}\\tSM:${name.split('_')[0..-2].join('_')}\\tPL:ILLUMINA\\tLB:${name}\\tPU:1\\tCN:${params.seq_center}\'" } + score = params.bwa_min_score ? "-T ${params.bwa_min_score}" : '' """ bwa mem \\ -t $task.cpus \\ -M \\ -R $rg \\ + $score \\ ${index}/${bwa_base} \\ $reads \\ | samtools view -@ $task.cpus -b -h -F 0x0100 -O BAM -o ${prefix}.bam - diff --git a/nextflow.config b/nextflow.config index 4a2e03ce..6ebe6d08 100644 --- a/nextflow.config +++ b/nextflow.config @@ -30,6 +30,7 @@ params { save_trimmed = false // Options: Alignments + bwa_min_score = false keep_mito = false keep_dups = false keep_multi_map = false From 328bb7dc2b0ce8be582c8658cc762f9b86c7c4d6 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 1 Jul 2020 14:51:22 +0100 Subject: [PATCH 222/226] Fix markdownlint --- docs/usage.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/docs/usage.md b/docs/usage.md index f772ef50..247b52a0 100755 --- a/docs/usage.md +++ b/docs/usage.md @@ -41,7 +41,7 @@ * [`--narrow_peak`](#--narrow_peak) * [`--broad_cutoff`](#--broad_cutoff) * [`--macs_fdr`](#--macs_fdr) - * [`--macs_pvalue`](#--macs_pvalue) + * [`--macs_pvalue`](#--macs_pvalue) * [`--min_reps_consensus`](#--min_reps_consensus) * [`--save_macs_pileup`](#--save_macs_pileup) * [`--skip_peak_qc`](#--skip_peak_qc) From a7e069b39dbf49230ba80c56b183ec3432cccfc7 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 1 Jul 2020 15:43:09 +0100 Subject: [PATCH 223/226] Change full test genome to hg19 --- conf/test_full.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/conf/test_full.config b/conf/test_full.config index e380e9c9..480e040f 100755 --- a/conf/test_full.config +++ b/conf/test_full.config @@ -15,5 +15,5 @@ params { input = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/design_full.csv' // Genome references - genome = 'hg38' + genome = 'hg19' } From 49e80e38773fb05eb7358a8205abe9602daa54f3 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 1 Jul 2020 16:05:52 +0100 Subject: [PATCH 224/226] Add params to schema --- nextflow_schema.json | 15 +++++++++++++++ 1 file changed, 15 insertions(+) diff --git a/nextflow_schema.json b/nextflow_schema.json index 0a570a76..149930a5 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -210,6 +210,11 @@ "description": "Reads mapping to multiple locations are not filtered from alignments.", "fa_icon": "fas fa-cart-arrow-down" }, + "bwa_min_score": { + "type": "integer", + "description": "Don\u2019t output BWA MEM alignments with score lower than this parameter.", + "fa_icon": "fas fa-hand-paper" + }, "skip_merge_replicates": { "type": "boolean", "default": false, @@ -259,6 +264,16 @@ "description": "Specifies broad cutoff value for MACS2. Only used when --narrow_peak isnt specified.", "fa_icon": "fas fa-hand-scissors" }, + "macs_fdr": { + "type": "number", + "description": "Minimum FDR (q-value) cutoff for peak detection, --macs_fdr and --macs_pvalue are mutually exclusive.", + "fa_icon": "fas fa-sort-amount-down" + }, + "macs_pvalue": { + "type": "number", + "description": "p-value cutoff for peak detection, --macs_fdr and --macs_pvalue are mutually exclusive. If --macs_pvalue cutoff is set, q-value will not be calculated and reported as -1 in the final .xls file.", + "fa_icon": "fas fa-sort-amount-down" + }, "min_reps_consensus": { "type": "integer", "default": 1, From 1c395ff3f4f79bc077a094a9ba7791233d5274d0 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 1 Jul 2020 23:09:42 +0100 Subject: [PATCH 225/226] Bump versions --- .github/workflows/ci.yml | 8 ++++---- CHANGELOG.md | 2 +- Dockerfile | 4 ++-- environment.yml | 2 +- nextflow.config | 4 ++-- 5 files changed, 10 insertions(+), 10 deletions(-) diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml index 675965f5..8a37ba68 100755 --- a/.github/workflows/ci.yml +++ b/.github/workflows/ci.yml @@ -35,13 +35,13 @@ jobs: - name: Build new docker image if: env.GIT_DIFF - run: docker build --no-cache . -t nfcore/atacseq:dev + run: docker build --no-cache . -t nfcore/atacseq:1.2.0 - name: Pull docker image if: ${{ !env.GIT_DIFF }} run: | docker pull nfcore/atacseq:dev - docker tag nfcore/atacseq:dev nfcore/atacseq:dev + docker tag nfcore/atacseq:dev nfcore/atacseq:1.2.0 - name: Install Nextflow run: | @@ -75,13 +75,13 @@ jobs: - name: Build new docker image if: env.GIT_DIFF - run: docker build --no-cache . -t nfcore/atacseq:dev + run: docker build --no-cache . -t nfcore/atacseq:1.2.0 - name: Pull docker image if: ${{ !env.GIT_DIFF }} run: | docker pull nfcore/atacseq:dev - docker tag nfcore/atacseq:dev nfcore/atacseq:dev + docker tag nfcore/atacseq:dev nfcore/atacseq:1.2.0 - name: Install Nextflow run: | diff --git a/CHANGELOG.md b/CHANGELOG.md index 36a10e05..bf2aad24 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -3,7 +3,7 @@ The format is based on [Keep a Changelog](http://keepachangelog.com/en/1.0.0/) and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0.html). -## [1.2.0] - 2020-06-04 +## [1.2.0] - 2020-07-02 ### `Added` diff --git a/Dockerfile b/Dockerfile index ecae8934..715e6892 100644 --- a/Dockerfile +++ b/Dockerfile @@ -7,10 +7,10 @@ COPY environment.yml / RUN conda env create -f /environment.yml && conda clean -a # Add conda installation dir to PATH (instead of doing 'conda activate') -ENV PATH /opt/conda/envs/nf-core-atacseq-1.1.1dev/bin:$PATH +ENV PATH /opt/conda/envs/nf-core-atacseq-1.2.0/bin:$PATH # Dump the details of the installed packages to a file for posterity -RUN conda env export --name nf-core-atacseq-1.1.1dev > nf-core-atacseq-1.1.1dev.yml +RUN conda env export --name nf-core-atacseq-1.2.0 > nf-core-atacseq-1.2.0.yml # Instruct R processes to use these empty files instead of clashing with a local version RUN touch .Rprofile diff --git a/environment.yml b/environment.yml index cadc2140..1fde3ddf 100755 --- a/environment.yml +++ b/environment.yml @@ -1,6 +1,6 @@ # You can use this file to create a conda environment for this pipeline: # conda env create -f environment.yml -name: nf-core-atacseq-1.1.1dev +name: nf-core-atacseq-1.2.0 channels: - conda-forge - bioconda diff --git a/nextflow.config b/nextflow.config index 6ebe6d08..5c08ccbf 100644 --- a/nextflow.config +++ b/nextflow.config @@ -99,7 +99,7 @@ params { // Container slug. Stable releases should specify release tag! // Developmental code should specify :dev -process.container = 'nfcore/atacseq:dev' +process.container = 'nfcore/atacseq:1.2.0' // Load base.config by default for all pipelines includeConfig 'conf/base.config' @@ -172,7 +172,7 @@ manifest { description = 'ATACSeq peak-calling and differential analysis pipeline.' mainScript = 'main.nf' nextflowVersion = '>=19.10.0' - version = '1.1.1dev' + version = '1.2.0' } // Function to ensure that resource requirements don't go beyond From a4751e91f4aa42873b78bc8912b53afcd0318259 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 1 Jul 2020 23:26:40 +0100 Subject: [PATCH 226/226] Update CHANGELOG --- CHANGELOG.md | 47 +++++++++++++++++++++++------------------------ 1 file changed, 23 insertions(+), 24 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index bf2aad24..bf724afa 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -8,14 +8,12 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Added` * [#63](https://github.com/nf-core/atacseq/issues/63) - Added multicore support for Trim Galore! -* [#71](https://github.com/nf-core/atacseq/issues/71) - consensus_peaks.mLb.clN.boolean.intersect.plot.pdf not generated * [#75](https://github.com/nf-core/atacseq/issues/75) - Include gene annotation versions in multiqc report * [#76](https://github.com/nf-core/atacseq/issues/76) - featureCounts coupled to DESeq2 * [#79](https://github.com/nf-core/atacseq/issues/79) - Parallelize DESeq2 * [#80](https://github.com/nf-core/atacseq/pull/80) - Added social preview image * [#97](https://github.com/nf-core/atacseq/issues/97) - PBC1, PBC2 from pipeline? * [#107](https://github.com/nf-core/atacseq/issues/107) - Add options to change MACS2 parameters -* [#109](https://github.com/nf-core/atacseq/issues/109) - Specify custom gtf but gene bed is not generated from that gtf? * [nf-core/chipseq#153](https://github.com/nf-core/chipseq/issues/153) - Add plotHeatmap * [nf-core/chipseq#159](https://github.com/nf-core/chipseq/issues/159) - expose bwa mem -T parameter * Regenerated screenshots and added collapsible sections for output files in `docs/output.md` @@ -34,10 +32,12 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Fixed` +* [#71](https://github.com/nf-core/atacseq/issues/71) - consensus_peaks.mLb.clN.boolean.intersect.plot.pdf not generated * [#73](https://github.com/nf-core/atacseq/issues/73) - macs_annotatePeaks.mLb.clN.summary.txt file is not created * [#86](https://github.com/nf-core/atacseq/issues/86) - bug in the plot_homer_annotatepeaks.r script * [#102](https://github.com/nf-core/atacseq/issues/102) - Incorrect Group ID assigned by featurecounts_deseq2.r * [#110](https://github.com/nf-core/atacseq/pull/110) - updated AWS test GitHub actions +* [#109](https://github.com/nf-core/atacseq/issues/109) - Specify custom gtf but gene bed is not generated from that gtf? * [nf-core/chipseq#118](https://github.com/nf-core/chipseq/issues/118) - Running on with SGE * [nf-core/chipseq#132](https://github.com/nf-core/chipseq/issues/132) - BigWig Error: sort: cannot create temporary file in '': Read-only file system * [nf-core/chipseq#154](https://github.com/nf-core/chipseq/issues/154) - computeMatrix.val.mat.gz files not zipped @@ -45,39 +45,38 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Dependencies` -* Add python `3.7.6` +* Add bioconductor-biocparallel `1.20.0` * Add markdown `3.2.2` -* Add pymdown-extensions `7.1` -* Add pygments `2.6.1` * Add pigz `2.3.4` -* Add r-tidyr `1.1.0` +* Add pygments `2.6.1` +* Add pymdown-extensions `7.1` +* Add python `3.7.6` * Add r-reshape2 `1.4.4` -* Add bioconductor-biocparallel `1.20.0` +* Add r-tidyr `1.1.0` +* Update ataqv `1.0.0` -> `1.1.1` +* Update bedtools `2.27.1` -> `2.29.2` +* Update bioconductor-deseq2 `1.20.0` -> `1.26.0` +* Update bioconductor-vsn `3.46.0` -> `3.54.0` +* Update deeptools `3.2.1` -> `3.4.3` +* Update fastqc `0.11.8` -> `0.11.9` * Update gawk `4.2.1` -> `5.1.0` +* Update homer `4.9.1` -> `4.11` +* Update macs2 `2.1.2` -> `2.2.7.1` +* Update multiqc `1.7` -> `1.8` +* Update picard `2.19.0` -> `2.23.1` +* Update pysam `0.15.2` -> `0.15.3` * Update r-base `3.4.1` -> `3.6.2` -* Update r-optparse `1.6.0` -> `1.6.6` * Update r-ggplot2 `3.1.0` -> `3.3.2` -* Update r-pheatmap `1.0.10` -> `1.0.12` * Update r-lattice `0.20_35` -> `0.20_41` +* Update r-optparse `1.6.0` -> `1.6.6` +* Update r-pheatmap `1.0.10` -> `1.0.12` +* Update r-scales `1.0.0` -> `1.1.1` * Update r-upsetr `1.3.3` -> `1.4.0` -* Update r-scales `1.0.0` -> `1.1.1` * Update r-xfun `0.3` -> `0.15` -* Update fastqc `0.11.8` -> `0.11.9` -* Update trim-galore `0.5.0` -> `0.6.5` * Update samtools `1.9` -> `1.10` -* Update picard `2.19.0` -> `2.23.1` -* Update pysam `0.15.2` -> `0.15.3` -* Update bedtools `2.27.1` -> `2.29.2` -* Update ucsc-bedgraphtobigwig `377` -> `357` -* Update deeptools `3.2.1` -> `3.4.3` -* Update macs2 `2.1.2` -> `2.2.7.1` -* Update homer `4.9.1` -> `4.11` -* Update ataqv `1.0.0` -> `1.1.1` * Update subread `1.6.4` -> `2.0.1` -* Update multiqc `1.7` -> `1.8` -* Update bioconductor-deseq2 `1.20.0` -> `1.26.0` -* Update bioconductor-vsn `3.46.0` -> `3.54.0` -* Remove r-reshape2 `1.4.3` +* Update trim-galore `0.5.0` -> `0.6.5` +* Update ucsc-bedgraphtobigwig `377` -> `357` ## [1.1.0] - 2019-11-05

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