Soft and Hard Homogenization DNA/RNA Extractions #139-146

20190726 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:

• The first and second elutions were spun down in separate microcentrifuge tubes to compare the quantity and quality between the first and second elution.

Coral fragments were randomly chosen from different timepoint bags.

Fragment preparation start 9:19 end 9:29
Soft/Hard homogenization start 9:30 end 9:49 RNA DNA Extraction start 9:50 end 11:40

10 μl aliquots of DNA and 5 μl aliquots of RNA.

Qubit start 11:00 end 12:15

Qubit used to check DNA and RNA quantity (ng/μl).
TapeStation used to check RNA quality.

DNA Standard 1: 178.41 ng/μl
DNA Standard 2: 2043.83 ng/μl
RNA Standard 1: 417.05 ng/μl
RNA Standard 2: 11005.81 ng/μl

Timepoint	Species	Coral ID	Extraction ID	Homogenization	Elution	DNA Reading 1	DNA Reading 2	Average DNA ng/μl	RNA Reading 1	RNA Reading 2	Average RNA ng/μl	RIN
20181020	Pocillopora	2564	139-1	soft	1	107	107	107	133	133	133	6.7
20181020	Pocillopora	2564	139-2	soft	2	34.2	34	34.1	11.2	11.2	11.2	6.8
20181020	Pocillopora	2564	140-1	hard	1	76.4	76	76.2	88.8	89	88.9	7.5
20181020	Pocillopora	2564	140-2	hard	2	22	22	22	14.8	14.8	14.8	7.8
20181103	Montipora	1467	141-1	soft	1	46.4	46.2	46.3	31.6	31.6	31.6	8.8
20181103	Montipora	1467	141-2	soft	2	9.72	9.68	9.7	**	**	**	**
20181103	Montipora	1467	142-1	hard	1	31.8	31.6	31.7	16.2	16.2	16.2	9.2
20181103	Montipora	1467	142-2	hard	2	9.9	9.74	9.82	**	**	**	**
20180923	Pocillopora	1303	143-1	soft	1	195	194	194.5	206	206	206	7.5
20180923	Pocillopora	1303	143-2	soft	2	33.6	33.6	33.6	29.2	29.2	29.2	7.1
20180923	Pocillopora	1303	144-1	hard	1	98.4	98	98.2	113	113	113	7.7
20180923	Pocillopora	1303	144-2	hard	2	18.9	18.8	18.85	18.4	18.4	18.4	6.9
20181117	Montipora	1204	145-1	soft	1	13.8	13.7	13.75	21.5	21.6	21.55	9
20181117	Montipora	1204	145-2	soft	2	11.6	11.6	11.6	**	**	**	**
20181117	Montipora	1204	146-1	hard	1	6	5.96	5.98	14.4	14.4	14.4	9.4
20181117	Montipora	1204	146-2	hard	2	8.14	8.06	8.1	**	**	**	**

Link to 20190726 TapeStation report, Extractions #139-146

Gel Electrophoresis Results: