INTRO
Steven asked that I isolate gDNA from a Pacific cod blood samples (GitHub Issue). Laura Spencer identified a set of samples (GitHub Issue). Below is the list of samples I isolated DNA from:
4B11B30B394770
The following blood samples were not located in the freezer boxes:
12B18B50B59B
METHODS
DNA Isolations
DNA was isolated using the Quick-DNA Miniprep Kit (ZymoResearch), according to the manufacturer’s protocol - including the recommended addition of beta-mercaptoethanol to the buffer. Pacific cod blood had been collected and frozen/stored at -80oC, with no preservatives/anticoagulants (e.g. heparin). Blood was thawed on ice and briefly centrifuged (20s @ 10,000g) to collect blood at bottom of tubes. Since the volumes of all blood samples was < 50uL, I added 200uL of g-DNA Lysis Buffer to each tube (per the manufacturer’s protocol for blood isolations). DNA was eventually eluted with 50uL of Elution Buffer.
DNA Quantification
Subsequently, DNA was quantified using the 1x dsDNA Broad Range Qubit Assay (ThermoFisher). One microliter of sample was combined with 199uL of the dye/buffer mix and measured on a Qubit 3.0.
DNA was stored in the same boxes and positions in which the blood had been stored in the -80oC freezer.
RESULTS
Qubit data
| sample_ID | Original sample conc.(ng/uL) |
|---|---|
| 4B | 9.36 |
| 11B | 212 |
| 30B | 1100 |
| 39B | 506 |
| 47B | 3.3 |
| 70B | 830 |