Soft and Hard Homogenization Extractions 665-674

20191208 E.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: X ng/μl
DNA Standard 2: X ng/μl
RNA Standard 1: X ng/μl
RNA Standard 2: X 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 1204 665 soft 35 34.8 34.9 30.6 30.8 30.7 8.3
20181117 Montipora 1204 666 hard 18.9 18.8 18.85 10.7 10.7 10.7 NA
20180922 Montipora 1609 667 soft 26 25.8 25.9 18.9 18.9 18.9 8.4
20180922 Montipora 1609 668 hard 12.5 12.4 12.45 10.2 10.2 10.2 NA
20180922 Pocillopora 2534 669 soft 105 104 104.5 108 108 108 7.4
20180922 Pocillopora 2534 670 hard 48.8 48.6 48.7 50.6 50.6 50.6 NA
20190112 Montipora 2403 671 soft 34.6 34.4 34.5 31.8 31.8 31.8 8.9
20190112 Montipora 2403 672 hard 20.8 20.8 20.8 13.2 13.2 13.2 NA
20181020 Montipora 2067 673 soft 16.5 16.4 16.45 5.02 5.08 5.05 **
20181020 Montipora 2067 674 hard 9.74 9.68 9.71 4 4 4 NA

Link to 20191209 TapeStation report, Extractions #665-674

Gel Electrophoresis Results:

20191208 Extractions #665-674

Written on December 19, 2019