Last time, I went over my plan to extract DNA from 4 random samples from SE AK Tanner crab, collected in 2005. This year was chosen because it’s the earliest samples we have - therefore, if they’ve got good DNA, all should have good DNA.
Plan was to pellet the hemolymph and then treat as a tissue sample. Samples were randomly chosen and numbered 1-4. A Qiagen DNEasy Blood + Tissue kit was used, followed by running them on the Qubit.
Important notes on use of Qiagen kit:
- Incubated for ~40 min with Protease K (until all was lysed)
- In the final elution step, used only 50 uL rather than 200 uL, as Shanelle mentioned that she had success with previous Tanner crab samples following that method
- Samples are currently stored in -20 (plate labeled with my name and date, samples labeled 1, 2, and 4, as 3 was discarded)
Important notes on use of Qubit:
- Ran both standards first
- Used 2 uL of sample (post-extraction of course), 198 uL of buffer
Alright, final concentrations as determined by Qubit:
Sample 1: 14.9 ng/ul
Sample 2: 2.18 ng/ul
Sample 3: failed to pellet
Sample 4: 15.3 ng/ul
These results are now available in the GitHub repo, just check out here
BioAnalyzer results are available within the same directory, here’s a link to that too!