Continuing Through-put Mo'orea Coral Extractions Four

One more 20 DNA only extractions from the first set of Porites and Pocillopora Corals from Mo’orea

Using the Zymo Quick-DNA Miniprep Plus kit

Sample Prep

Sample # Type
245 Pocillopora verrucosa
174 Pocillopora verrucosa
184 Pocillopora verrucosa
224 Pocillopora verrucosa
39 Massive Porites
38 Pocillopora verrucosa
210 Massive Porites
278 Massive Porites
314 Massive Porites
272 Massive Porites
284 Massive Porites
160 Pocillopora verrucosa
196 Massive Porites
35 Pocillopora verrucosa
52 Massive Porites
53 Massive Porites
36 Massive Porites
237 Pocillopora verrucosa
239 Pocillopora verrucosa
51 Massive Porites
  • 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. 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

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
245 192 19503 49 48 48.5
174 192 19503 61 60.6 60.8
184 192 19503 47.2 47.2 47.2
224 192 19503 31.6 31.4 31.5
39 192 19503 9.63 9.14 9.35
38 192 19503 20.6 20.4 20.5
210 192 19503 4.94 4.74 4.84
278 192 19503 30.2 29.8 30
314 192 19503 13.3 13.2 13.2
272 192 19503 7.82 7.62 7.72
284 192 19503 11.3 11.2 11.2
160 192 19503 38.2 38 38.1
196 192 19503 26.6 26.4 26.5
35 192 19503 68.6 68 68.3
52 192 19503 7.58 7.48 7.53
53 192 19503 19.2 19.2 19.2
36 192 19503 19.5 19.6 19.5
237 192 19503 125 123 124
239 192 19503 90.4 89.2 89.8
51 192 19503 39.8 39.6 39.7

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

gel

Written on April 29, 2019