INTRO
Steven asked that I isolate gDNA from a subset of lake trout (S.namaycush) liver samples (GitHub Issue) to use for targeted DNA methylation analyses via qPCR.
METHODS
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. Liver tissues had been stored @ -20oC in RNALater (Ambion). I thawed the tubes, transferred the entire liver tissue (which was very small) to a disposable mortar/pestle combo (1.5mL tube) and homogenized. DNA was eventually eluted with 50uL of Elution Buffer.
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 a box labelled “Lake Trout Liver gDNA” in the FTR 209 -20oC freezer.
RESULTS
Qubit data
| sample_ID | Original sample conc. (ng/uL) |
|---|---|
| FA066 | 340 |
| FA067 | 760 |
| FA068 | 548 |
| FA069 | 202 |
| FA070 | 454 |
| FA077 | 532 |
| FA078 | 236 |
| FA079 | 318 |
| FA080 | 120 |
| FA081 | 474 |