Another 20 DNA only extractions from Porites and Pocillopora Corals from Mo’orea

Using the Zymo Quick-DNA Miniprep Plus kit

Sample Prep

Sample #	Type
42	Pocillopora verrucosa
46	Pocillopora verrucosa
60	Pocillopora verrucosa
161	Pocillopora verrucosa
165	Pocillopora verrucosa
169	Pocillopora verrucosa
182	Pocillopora verrucosa
183	Pocillopora verrucosa
188	Pocillopora verrucosa
202	Massive Porites
204	Massive Porites
214	Massive Porites
218	Massive Porites
226	Massive Porites
236	Pocillopora verrucosa
243	Pocillopora verrucosa
270	Massive Porites
307	Massive Porites
310	Massive Porites
311	Massive Porites

• New Beads were poured into sample tubes. The new beads should be easier to pipette the liquid out of them as the do not get sucked up by the p20
• Samples were homogenized by vortexing for ~60 seconds for all samples
• Most of the liquid from the tubes was removed by pipetting. I guessed that this was about 420µl. The tubes contained a small amount of liquid and un-homogenized tissue left, so 200µl of DNA/RNA shield was added to each tube and those were put back into the -20
• Following recommendations for samples in DNA/RNA Shield from the kit protocol, 210µl of Solid Tissue Buffer and 14µl of Proteinase K were added to each sample
• Samples were votexed, spun down, and incubated at 55 degrees C for 5 hours shaking at 600rpm

Following DNA extraction went along exactly as previous post

6µl of sample were aliquoted out for Qubit and Gel the next day to avoid thawing the stock.

Qubit

• Broad Range dsDNA Qubit protocol
• The Qubit was performed by Marygrace Trusdell
• All samples were read twice

Sample	DNA Standard 1 (RFU)	DNA Standard 2 (RFU)	DNA 1 (ng/µl)	DNA 2 (ng/µl)	Average DNA
42	196	16886	36.8	37.2	37
46	196	16886	52.8	51.8	51.3
60	196	16886	65.2	67.6	66.4
161	196	16886	52.8	54.2	53.5
165	196	16886	62.4	61.4	61.9
169	196	16886	40.2	38.6	39.4
182	196	16886	79.2	78.2	78.7
183	196	16886	47.6	48.8	48.2
188	196	16886	111	110	110.5
202	196	16886	15.2	14.8	15
204	196	16886	43.6	44	43.8
214	196	16886	48.4	46.8	47.7
218	196	16886	29.6	28.8	29.3
226	196	16886	18.3	17.8	18
236	196	16886	45.4	43.6	44.5
243	196	16886	77.2	76.4	76.7
270	196	16886	18.7	18.6	18.6
307	196	16886	13.7	12.8	13.4
310	196	16886	23	22.6	22.8
311	196	16886	17.4	17.4	17.4

67 and 220 were re-dos, so there might not have been that much left in the first place. I need to do a gel to see how the quality is.

Gel Verification

• A 1.5% agarose gel was ran to check the integrity of the genomic DNA
• Following the PPP Lab protocol
• Samples 67 and 220 were not run on the gel