Continuing extracting RNA and DNA from settled Mcap spat on plugs, Hawaii - 092425

Extracting RNA and DNA from spat on plugs Montipora capitata for the Hawaii settlement experiment

Protocols used

I followed the extraction protocol linked above exactly, final elution volume for both RNA and DNA samples is 15uL. The plugs were flash frozen and stored in whirlpacks. These are the pictures of the plugs before sampling:

![B6before.jpg](https://github.com/FScucchia-LabNotebooks/FScucchia_Putnam_Lab_Notebook/blob/master/images/B6_before.jpg?raw=true) B11_before.jpg A6_before.jpg C6_before.jpg D6_before.jpg D11_before.jpg D12_before.jpg A4_before.jpg _Note: I had already extracted sample A4, I’m trying again to see if I can get a better RIN.

I took microbiome samples as well, using sterile razor blades, scraping off the surface away from the spat, one sample per plug. These microbiomes samples were named with the sample name and an “X”. These are the pictures of the plugs after sampling:

B6_after.jpg B11_after.jpg A6_after.jpg C6_after.jpg D6_after.jpg D11_after.jpg D12_after.jpg A4_after.jpg

This is how the samples in shield looked like before extraction: samples_shield4.jpg samples_shield5.jpg

Qubit Results

I used the high sensitivity RNA Qubit Protocol linked 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: 47.89
  • Standard 2: 919.85
Scraping date   Extraction date   sample_id   Species   Treatment   RNA_read1   RNA_read2   RNA_AVG (ng/ul)
20250924   20250924   B6   Montipora capitata   SNAP-TAG treatment   59.4   58.6   59.0
20250924   20250924   B11   Montipora capitata   SNAP-TAG treatment   49.0   47.4   48.2
20250924   20250924   A6   Montipora capitata   SNAP-TAG control   46.8   46.0   46.4
20250924   20250924   C6   Montipora capitata   SNAP-TAG control   27.0   27.0   27.0
20250924   20250924   D6   Montipora capitata   SNAP-TAG treatment   61.0   60.6   60.8
20250924   20250924   D11   Montipora capitata   SNAP-TAG treatment   36.8   36.6   36.7
20250924   20250924   D12   Montipora capitata   SNAP-TAG control   30.4   30.4   30.4
20250924   20250924   A4   Montipora capitata   SLIPS treatment   77.8   76.0   76.9

Tapestation Results tapeStatio_McapPlug4.png

Written on September 24, 2025