Sam’s Notebook: DNA Isolation, Quantification, and Gel – C.bairdi gDNA Sample 6129_403_26

In order to do some genome sequencing on C.bairid and Hematodinium, we need hihg molecular weight gDNA. I attempted this twice before, using two different methods (Quick DNA/RNA Microprep Kit (ZymoResearch) on 20200122 and the E.Z.N.A Mollusc DNA Kit (Omega) on 20200108) using ~10yr old ethanol-preserved tissue provided by Pam Jensen. Both methods yielded highly degrade gDNA. So, I’m now attempting to get higher quality gDNA from the RNAlater-preserved hemolymph pellets from this experiment.

Used the Quick DNA/RNA Microprep Kit (ZymoResearch) to perform four parallel isolations using four separate aliquots of sample 6129_403_26 with the following changes/notes:

  • Used 75uL of each hemolyph pellet slurry from each tube
  • Eluted all columns with a single 30uL elution (i.e. used 30uL for first column, then used that same elution to elute the next column, etc.) to try to concentrate the sample.

DNA was quantified using the Roberts Lab Qubit 3.0, using 1uL of sample, with the ds DNA BR Assay (Invitrogen).

Used 4uL of sample (~200ng) to run on 1x low-TAE 0.8% agarose gel.