Objective and summary:
-
REDO complete extraction and qubit quantification using ** ethanol-fixed adductor muscle tissue of target scallops**
- 60 mg of tissue
- 3 hr Proteinase K incubation at 120 RPM and 55C - vortexed every hour
- doubled the volume of TL Buffer, HBC buffer and 100% ethanol for the 60 mg samples (did not double proteinase K)
- of the 18 target F0 adult scallops extracted (all spawned 7/26/22) =, 17 were extracted again to attempt to yield higher DNA yield (all but ID#9)
- DNA was extracted using the OMEGA EZNA Tissue kit protocol here
- Ran Qubit dsDNA BR kit following standard protocol thermosci online pdf
Qubit Results
| Sample | DNA (ng/uL) | |
|---|---|---|
| RUN 1 | RUN 2 | |
| Standard | 67.39 | 68.57 |
| Standard | 7900.23 | 7780.44 |
| 1 | 16.5 | 16.6 |
| 2 | 5.67 | 5.52 |
| 3 | 20.4 | 19.7 |
| 4 | 6.14 | 5.97 |
| 5 | 5.57 | 5.18 |
| 7 | 5.52 | 5.32 |
| 11 | 9.91 | 9.57 |
| 17 | 9.86 | 9.63 |
| 18 | 12.8 | 12.0 |
| 19 | 5.51 | 5.25 |
| 20 | 5.08 | 5.04 |
| 21 | 6.44 | 6.16 |
| 23 | 17.7 | 17.2 |
| 24 | 2.54 | 2.49 |
| 25 | 5.47 | 5.41 |
| 26 | 5.74 | 5.64 |
| 27 | 3.97 | 3.70 |
Check previous Qubit notebook posts to see if what I did today increased our yield. Oddly there is not an expected high difference (i.e. upwards of 40-80 ng/ul in previous test) but I can somewhat confirm now that this the nature of our samples.
moving forward with lcWGS library prep? we need 2.5 ng/ul DNA to start Nina’s protocol, this Qubit run looks OK