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

Protocols used

• Dual RNA/DNA extraction micro kit
• QBIT protocol
• Gel protocol

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:

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:

This is how the samples in shield looked like before extraction:

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: 57.48
• Standard 2: 1088.70

Tapestation Results

Gel

• I used the protocol linked above with few modifications. I ran 3.5ul of RNA and 3.5ul of DNA. I added 4ul of gel green to the gel.