Soft and Hard Homogenization Extractions 645-652

20191205 M.S. DNA/RNA Extractions from Montipora capitata and Pocillopora acuta adult coral fragments from Holobiont Integration Hawaii 2018 project.

Soft and Hard homogenization, and DNA/RNA Extractions followed this protocol. General Zymo Duet DNA/RNA Extraction protocol found here. With the following changes:

  • DNA: The first and second elutions were spun down in separate microcentrifuge tubes to compare the quantity and quality between the first and second elution. The first elution had no incubation period (previously 5 minutes) with 10 μl of Tris buffer and the second elution had a 15 minute incubation period (previously 5 minutes) with 100 μl of Tris buffer.
  • 10 μl aliquots of DNA and 5 μl aliquots of RNA (instead of 10 μl of each).

Qubit used to check DNA and RNA quantity (ng/μl).
TapeStation used to check RNA quality.

High Sensitivity:
DNA Standard 1: 47.64 ng/μl
DNA Standard 2: 25,437.24 ng/μl
RNA Standard 1: 52.52 ng/μl
RNA Standard 2: 1,057.21 ng/μl

Timepoint Species Coral ID Extraction ID Homogenization DNA Reading 1 DNA Reading 2 Average DNA ng/μl RNA Reading 1 RNA Reading 2 Average RNA ng/μl RIN
20181117 Montipora 1332 645 soft 43.4 43.2 43.3 19.9 19.9 19.9 7.9
20181117 Montipora 1332 646 hard 22.2 22.2 22.2 9.4 9.44 9.42 NA
20180929 Montipora 1212 647 soft 34 34 34 22.4 22.4 22.4 8.4
20180929 Montipora 1212 648 hard 24.2 24.2 24.2 14.2 14.1 14.15 NA
20181020 Montipora 3753 649 soft 46 46 46 28.6 28.6 28.6 8.9
20181020 Montipora 3753 650 hard 17.9 17.9 17.9 11 11 11 NA
20180922 Montipora 1481 651 soft 40.6 40.4 40.5 24 24 24 8.2
20180922 Montipora 1481 652 hard 19.7 19.6 19.65 11.8 11.8 11.8 NA

Link to 20191205 TapeStation report, Extractions #645-652

Gel Electrophoresis Results:

20191205 Extractions #645-652

Written on December 5, 2019