Starting in on RNA extractions of samples from Day 12 (controls) and Day 6 (exposed and controls) from the multi-species experiment. See post for details.
Samples Chosen
| SampleID | Species | Bin | ExperimentDay | Experiment |
|---|---|---|---|---|
| 423 | pycnopodia_helianthoides | 1 | 6 | exposed |
| 426 | pycnopodia_helianthoides | 9 | 6 | control |
| 430 | dermasterias_imbricata | 10 | 6 | control |
| 434 | pisaster_ochraceus | 3 | 6 | exposed |
| 438 | pycnopodia_helianthoides | 11 | 6 | control |
| 441 | pycnopodia_helianthoides | 4 | 6 | exposed |
| 452 | pisaster_ochraceus | 6 | 6 | exposed |
| 454 | dermasterias_imbricata | 14 | 6 | control |
| 464 | pisaster_ochraceus | 8 | 6 | exposed |
| 534 | pycnopodia_helianthoides | 11 | 12 | control |
| 550 | dermasterias_imbricata | 14 | 12 | control |
| 563 | pisaster_ochraceus | 16 | 12 | control |
RNA Extraction
Notes:
- the Kits i have are from 2024 – this is 1 year, which from what I can find online - they are still usable
- DNAse I –> from 2024 and was reconstituted then and stored at -20C since then (Cat: E1009-A; Lot: 223372)
- Samples 434 and 534 have brownish tissue at the bottom (the pellet + 300ul DNA/RNA Shield - prior to starting the extraction protocol)
- Proteinase K aliquot from -20C freezer had pelleted after thawing, so I vortexed
Start at 10:15am (take samples from freezer and thaw on wet ice)
Start extraction protocol at: 10:40am
Start incubation at 10:45am, continue with extraction at 11:25am
Finished whole protocol at 12:59pm
Put extracted RNA in -80C immediately.
DNA (on yellow filters) is in -20C (needs to finish extraction process if want DNA).
To Do Soon:
Run these 12 samples on Qubit.
Continue on through extractions.