RNA extraction and qPCR for snap frozen Lifestage Carryover samples

[AHuffmyer]

RNA extractions and qPCR for Lifestage Carryover project for 60 snap frozen samples.

These can be done in the same way as the previous round for the same targets (ATP synthase, Viperin, HSP70, HSP90, Citrate Synthase, cGAS, DMNT1 normalized to GADPH).

In the last round of analyses, we were not able to get RNA from some samples, leading to low sample size. This task is to conduct the extractions and qPCR on snap frozen samples to supplement sample size.

The adult group had sufficient sample sizes (n=8 per group), so here we will only use samples for spat, seed, and juveniles.

Note that the spat and seed samples are of whole individuals and will need to be crushed/homogenized during extractions.

The sample list can be found here. Please refer to the "notes" column for clarification on sample/tube IDs and freezer location.

I have attached pictures of the two boxes that contain all samples. Please refer to the spreadsheet linked above for the tube ID's to use. The boxes also contain "adult" samples that will not be processed. Tubes in the boxes are arranged in numerical order.

@kubu4 - does this sound ok? Let me know if I can help with clarifying any questions. Are we good on supplies for processing these 60 samples?

Samples are in the two "LCO Phys" boxes in the image below in the middle freezer (top shelf).

[kubu4]

Are we good on supplies for processing these 60 samples

Nope. We'll need to order more reagents.

I'll get the necessary stuff ordered and will tackle once we start getting supplies in.

[AHuffmyer]

@kubu4 what is the status of supply orders for the remaining qPCR samples? Thank you!

[kubu4]

We have received all supplies.

This process might be getting "outsourced." Discussed this briefly with @sr320 , but nothing's been finalized yet.