Soft and Hard Homogenization Extractions 685-692

20191210 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.

Broad Range: DNA Standard 1: X ng/μl
DNA Standard 2: X ng/μl

High Sensitivity: 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
20190112 Montipora 2990 685 soft 28.4 28 28.2 16.6 16.6 16.6 8.3
20190112 Montipora 2990 686 hard 15.2 15 15.1 12.7 12.6 12.65 NA
20180923 Montipora 2986 687 soft 12 11.9 11.95 15.6 15.5 15.55 8.7
20180923 Montipora 2986 688 hard 10.4 10.3 10.35 8.32 8.24 8.28 NA
20180922 Montipora 2153 689 soft 37.2 36.6 36.9 38.2 38 38.1 9
20180922 Montipora 2153 690 hard 21.2 20.8 21 15.7 15.7 15.7 NA
20181103 Montipora 1467 691 soft 28.4 27.8 28.1 15.6 15.5 15.55 8.6
20181103 Montipora 1467 692 hard 12.9 12.6 12.75 8.62 8.54 8.58 NA

Link to 20191210 TapeStation report, Extractions #685-692

Gel Electrophoresis Results:

20191209 Extractions #685-692

Written on December 19, 2019