Sam’s Notebook: DNA Shearing & Bioanalyzer – Lotterhos C.virginica Mantle gDNA from 2018114

Crassostrea virginica mantle gDNA that had previously been isolated on 20181114 was evaluated for integrity via agaroase gel yesterday (20190305). Everything looked pretty good, so proceeded with shearing in preparation for MBD enrichment.

Aliquoted 2500ng of DNA in a final volume of 100uL of each sample in 0.5mL snap cap (polypropylene) tubes for shearing in the Seeb Lab sonicator (Diagenode Bioruptor 300) with the following settings:

  • 30 cycles
  • 30 seconds on
  • 30 seconds off

Cycle settings on Diagenode Bioruptor for DNA shearing

Calculations for DNA dilutions are here (Google Sheet):

Ideally, this will shear DNA to a target size around 300bp.

After shearing, ran all samples on the Bionalyzer 2100 (Agilent) using a DNA 12000 chip according to manufacturer’s protocol.