INTRO

Steven asked that I isolate gDNA from more Siscowet lake trout (S.namaycush) liver samples (GitHub Issue) to send to Angie for sequencing - part of Rick’s project.

METHODS

DNA was isolated using the Quick-DNA/RNA Miniprep Plus Kit (ZymoResearch), according to the manufacturer’s protocol. Liver tissues had been stored @ -20^oC 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 in 1x DNA/RNA Shield. Insoluble material was pelleted and the remainder of the protocol was followed (note: I did not perform Proteinase K lysis). DNA was eventually eluted with 50uL of water.

Subsequently, DNA was quantified using the 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.

All samples were subjected to concentrating using a SpeedVac on the “High Drying Rate” setting for ~1hr, as Angie needs highly concentrated DNA for sequencing, with a targeted final volume of ~10uL.

DNA was stored in a box labelled “Lake Trout Liver gDNA” in the FTR 209 -20^oC freezer.

RESULTS

Qubit data

20250708-snam-qubit-DNA.br-liver (Google Sheet)

sample_ID	Concentration (ng/uL)	Volume (uL)	Yield (ng)
FA 244	29	40	1160
FA 245	270	40	10800
FA 251	104	40	4160
FA 252	183	40	7320
FA 253	195	40	7800

Note

This table details sample info before concentrating. Samples were not measured after concentrating.