And 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
283	Massive Porites
306	Massive Porites
206	Massive Porites
277	Massive Porites
217	Pocillopora verrucosa
225	Pocillopora verrucosa
228	Massive Porites
274	Massive Porites
61	Pocillopora verrucosa
233	Massive Porites
285	Massive Porites
313	Massive Porites
240	Pocillopora verrucosa
57	Massive Porites
189	Pocillopora verrucosa
212	Massive Porites
281	Massive Porites
47	Pocillopora verrucosa
199	Massive Porites
190	Pocillopora verrucosa

• 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 400µl. 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, 200µl of Solid Tissue Buffer and 12µl 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
283	184	20179	13.6	13.6	13.6
306	184	20179	26.2	26.2	26.2
206	184	20179	27.6	27.6	27.6
277	184	20179	30.2	30.2	30.2
217	184	20179	76.6	76	76.3
225	184	20179	58.8	58	58.4
228	184	20179	15.5	15.3	15.4
274	184	20179	16.9	16.7	16.8
61	184	20179	46.8	46.4	46.6
233	184	20179	18.7	18.5	18.6
285	184	20179	8.33	8.34	8.34
313	184	20179	12.5	12.3	12.4
240	184	20179	87	86.2	86.6
57	184	20179	30.4	30.2	30.3
189	184	20179	58	57.2	57.6
212	184	20179	15	14.2	14.6
281	184	20179	39.4	38.8	39.1
47	184	20179	57.4	56	56.7
199	184	20179	72.8	71.4	72.1
190	184	20179	77	76.8	76.9

Gel Verification

• A 1.5% agarose gel was ran to check the integrity of the genomic DNA
• Following the PPP Lab protocol
• Gel was made and run by Marygrace Trusdell