Ronit’s Notebook: qPCR Prep/Spreadsheet Organization

In preparation for the RNA extraction from the heat stress C. Gigas samples, Sam and I went over some sample organization/prep. I updated my spreadsheet, which was linked in my previous notebook entry, so that it now has columns to show treatment, ploidy, and tissue type. Also–there are now entries for replicate samples as well (i.e. D01-C2), so that every sample tube has a corresponding entry in the spreadsheet.

I then chose 2 samples from each treatment to prep for RNA extraction and subsequent qPCR. As there were 8 distinct treatment groups, we pulled a total of 16 samples:

D01, D02 — Diploid oysters exposed to control conditions (water in aquarium)

D09, D10 — Diploid oysters exposed to control conditions (water in aquarium); subsequently exposed to 1 hr acute heat shock at 45 degrees Celsius.

D11, D12 — Diploid oysters exposed to desiccation + elevated temperature (27 degrees Celsius) for 24 hrs

D19, D20 — Diploid oysters exposed to desiccation + elevated temperature (27 degrees Celsius) for 24 hrs; subsequently exposed to 1 hr acute heat shock at 45 degrees Celsius.

T01, T02 — Triploid oysters exposed to control conditions (water in aquarium)

T09, T10 — Triploid oysters exposed to control conditions (water in aquarium); subsequently exposed to 1 hr acute heat shock at 45 degrees Celsius.

T11, T12 — Triploid oysters exposed to desiccation + elevated temperature (27 degrees Celsius) for 24 hrs

T19, T20 — Triploid oysters exposed to desiccation + elevated temperature (27 degrees Celsius) for 24 hrs; subsequently exposed to 1 hr acute heat shock at 45 degrees Celsius.

These tubes were pulled from the -80 freezer and kept in a new, unlabeled box (which is also in the -80 freezer and has been updated in the -80 Inventory Map). We then made sure that we had all the materials on hand for the RNA extraction next week.