Holobiont Integration March Extractions

March DNA RNA Extractions

20200226 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.

DNA Standard 1: 144.56 ng/μl
DNA Standard 2: 21,023.95 ng/μl
RNA Standard 1: 51.95 ng/μl
RNA Standard 2: 975.22 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
20191018 Montipora 1464 689 soft 28 27.8 27.9 35.2 35.2 35.2 8.7
20191018 Montipora 1464 690 hard 16.7 16.6 16.65 12.6 12.6 12.6 NA
20191018 Montipora 1532 691 soft 57.6 57.4 57.5 61.2 61.2 61.2 8.7
20191018 Montipora 1532 692 hard 29.2 29.2 29.2 29.2 29.2 29.2 NA
20191018 Montipora 1417 693 soft 39 38.8 38.9 23.6 23.8 23.7 8.9
20191018 Montipora 1417 694 hard 27.2 27 27.1 11.3 11.4 11.35 NA
20191018 Montipora 1546 695 soft 24.6 24.6 24.6 28.6 28.6 28.6 9
20191018 Montipora 1546 696 hard 22.8 22 22.4 22.4 22.4 22.4 NA
20191018 Montipora 1259 697 soft 40.8 40.6 40.7 38.6 38.6 38.6 9.2
20191018 Montipora 1259 698 hard 23.6 23.4 23.5 17.1 17 17.05 NA

Link to 20200226 TapeStation report, Extractions #689-698

Gel Electrophoresis Results:

20200226 Extractions #689-698

Written on February 27, 2020