After speaking with Axa regarding the DNA concentrations, he would like the DNA from the ethanol-fixed tissue to be more concentrated, and he wants them in ddH2O instead of Buffer AE (from the Qiagen DNeasy Kit). So, I preformed a standard ethanol precipitation. Added 0.1 volumes of 3M sodium acetate (pH = 5.2) [15uL], 2.5 volumes of 100% ethanol [412.5uL] and incubated @ -20C over the weekend.
Pelleted DNA by spinning 16,000g, 15mins @ 4C. Discarded supe, washed pellets with 1mL 70% ethanol and re-pelleted the DNA. Performed a second wash with 70% EtOH, pelleted DNA, discarded supe, resuspended DNA in 75uL of NanoPure H2O, and spec’d on NanoDrop1000.
Results:
Will spec when I re-isolate DNA from “fresh” geoduck samples for coordinating Axa’s DNA sequencing project with our potential RNA-seq project. See 20140219.