Soft and Hard Homogenization Extractions 675-684

20191209 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
20181215	Pocillopora	1147	675	soft	54	53.8	53.9	54.8	54.8	54.8	5.9
20181215	Pocillopora	1147	676	hard	19.5	19.5	19.5	19.8	19.8	19.8	NA
20180923	Montipora	1321	677	soft	37.4	37.2	37.3	35.2	35.2	35.2	9
20180923	Montipora	1321	678	hard	16.9	16.8	16.85	13.5	13.5	13.5	NA
20180929	Montipora	1604	679	soft	29.2	29	29.1	18.8	18.8	18.8	8.1
20180929	Montipora	1604	680	hard	30.6	30.4	30.5	13.4	13.4	13.4	NA
20181006	Montipora	1645	681	soft	30.4	30.2	30.3	23.8	23.8	23.8	8.9
20181006	Montipora	1645	682	hard	20.6	20.4	20.5	14.3	14.3	14.3	NA
20181020	Montipora	2067	683	soft	24.8	24.6	24.7	10.8	10.8	10.8	8.6
20181020	Montipora	2067	684	hard	13	13	13	5.9	5.84	5.87	NA