20191216 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: 164 ng/μl
DNA Standard 2: 23,368 ng/μl

High Sensitivity: RNA Standard 1: 50.53 ng/μl
RNA Standard 2: 1,061 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
20180922	Montipora	2734	693	soft	38.8	38.6	38.7	4.8	4.6	4.7
20180922	Montipora	2734	694	hard	21	21	21	**	**	**	NA

Re-extraction:

Broad Range: DNA Standard 1: 166 ng/μl
DNA Standard 2: 23,551 ng/μl

High Sensitivity: RNA Standard 1: 53.50 ng/μl
RNA Standard 2: 1,065 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
20180922	Montipora	2734	695	soft	74.2	73	73.6	54.6	54.2	54.4	8.3
20180922	Montipora	2734	696	hard	35.4	34	34.7	22.4	22.4	22.4	NA

Tubes labeled as extraction IDs 685, 686, 687, 688. The correct extraction IDs are 693, 694, 695, 696.

Link to 20191216 TapeStation report, Extractions #695

Gel Electrophoresis Results: