Continuing extracting RNA and DNA from Pacu, Mcap, Pcomp - Hawai TPC - 120424

Extracting RNA and DNA from fragments of Pocillopora acuta, Montipora capitata, Porites compressa for the Hawaii TPC project

Protocols used

I’m continuing extracting RNA and DNA from frozen fragments of Pacuta, Mcap and Pcomp using the Zymo Quick-DNA/RNA Miniprep Plus Kit and the protocol linked above. The kit protocol is found here.

  • I eluted RNA samples in 80ul and DNA samples in 80ul.

Qubit Results I used Broad range RNA Qubit Protocol inked above. I will check the DNA samples after finishing with all extractions (for all samples). RNA samples were read twice, standard only read once.

  • Standard 1: 435.27
  • Standard 2: 9432.82
Clipping date   Extraction date   colony_id   Species   Temp.Cat   RNA_read1   RNA_read2   RNA_AVG (ng/ul)
20241203   20241204   Pacu-B8   Pocillopora acuta   25   23.6   23.4   23.5
20241203   20241204   Mcap-B7   Montipora capitata   35   24.4   23.0   24.2
20241202   20241204   Pcomp-G1   Porites compressa   18   31.6   30.2   30.9
20241129   20241204   Mcap-G6   Montipora capitata   12   21.4   21.0   21.2
  • Standard 1: 469.49
  • Standard 2: 10098.72
Clipping date   Extraction date   colony_id   Species   Temp.Cat   RNA_read1   RNA_read2   RNA_AVG (ng/ul)
20241203   20241204   Pacu-E9   Pocillopora acuta   30   64.4   63.6   64.0
20241202   20241204   Mcap-G1   Montipora capitata   18   54.4   53.4   53.9
20241202   20241204   Pcom-E1   Porites compressa   18   18.8   18.6   18.7
20241203   20241204   Pcom-B7   Porites compressa   35   22.8   22.4   22.6

Gel

  • I used the protocol linked above with few modifications. I ran 11ul of RNA and 9ul of DNA. I added 4ul of gel green to the gel.

Gel_DNA_RNA_04Dec2024.png

Note: Looks like for McapG6 the DNAse tretment didn’t work well, I’m re-extracting this sample on the 16th

Written on December 4, 2024