See post for details on the second batch of RNA extractions for the new dataset.

Samples Chosen

Note: Like in the previous batch (Notebook post: here), I am randomizing the samples from Day 6 (exposeed + control) and Day 12 (control).

RNA Extraction

Notes:

Start labeling at 1:26pm
Start extraction protocol at 2:20pm; incubate 2:30-3:07pm
End at 4:55pm

• Added 96mL of 100% EToH to a new bottle (From the opened 2024 kit) of DNA/RNA Wash Buffer because I think there isn’t enough currently for these 18 samples –> didn’t end up using for this batch, but it’s ready for tomorrow

Qubit Results:

Total samples to run: 18+12 = 30
I’m going to do them in two batches.
So each batch will be: 15 samples + 2 standards + 1 extra rxn == 18 preps

I let the two standards come up to room temp (they’re stored in fridge).

Therefore:
Working solution:
18 x 1ul RNA HS Dye = 18ul RNA HS Dye
18 x 199ul RNA HS Buffer = 3582ul RNA HS Buffer

Standards:
10ul S1 + 190ul working Solution
10ul S2 + 190ul working Solution

Samples:
1ul Sample + 199ul working solution

Vortex all samples and standards after working solution added for 3-5mins.
Incubate at room temp for 2 minutes.

Results:

BATCH 1 (423-431)
S1 –> 68.35
S2 –> 971.17

BATCH 2 (433-564)
S1 –> 71.63
S2 –> 1120.38

LONG STORY SHORT:

THERE IS ENOUGH RNA IN ALL SAMPLES SO FAR FOR RNASEQ HUZZAH