Mon. Jan 28, 2019, Geoduck Broodstock DNA extraction from Hemocytes

Kaitlyn and I isolated DNA from four Jan. 4 hemocyte samples. Two samples were from Tank 2 (low pH) and two were from Tank 3 (ambient) treated animals. We used the EZNA kit and followed the manual with the following exceptions:

  • We started on step 2 because we had cells not tissue, so no tissue homogenization was necessary
  • Heat block in rm 213 was 59-65C instead of 60C because the digital setting was a few degrees off from the alcohol thermometer.
  • In step 6, the volumes transferred were:
    • 250uL for sample 16
    • 300uL for sample 15
    • 350uL for sample 26
    • 400uL for sample 25
  • We eluted with 2 x 50uL elution buffer warmed to 70C.

Then used 1 uL of each sample for Qubit BR.

Standard 2 read 98 ng/uL and 102 ng/uL, so average is 100 ng/uL which is what it should be.

Here are the following qubit readings for the samples:

Animal ID Tank Treatment Conc. (ng/ul) Total DNA (ng)
15 3 amb 12.3 1230
16 3 amb 56 5600
025 2 low (pH 6.8) 8.3 830
026 2 low (pH 6.8) 7.4 740

We will send these for WGBS.

Written on January 28, 2019