September 02, 2024

• First, I completed DNA extractions on the remaining 22 mussel mantle tissue samples based on the gel results from 8/30/24. The first run of extractions only completed 18 of the 22 samples because samples 106, 107, 269 and 272 didn’t make it past step 6 in the protocol. Each of the samples required an additional 45 minutes of incubation in the Proteinase K Solution to get to almost complete solubility, but didn’t yield the ‘upper aqueous phase’, the separation of working material and contaminants and inhibitors, in subsequent steps. Once I finished the remaining extractions, I went back and started those four from the beginning. The instructions state that 30 mg is the ideal tissue quantity for extraction, but give a range up to 50 mg of tissue. This didn’t prove a problem for any of the other samples which fell between 30 - 50 mg, but only 30 mg of tissue was dissected out for extraction from each of these samples and that worked in completing the process.
  
• Next I checked concentration on all 22 samples using the Quibit HS DNA assay. All sample concentrations were above 50 ng/uL so I recorded the concentrations and moved into running a gel.
  
• Finally, I ran a 1% agarose gel to do a preliminary check for DNA integrity. That ran for 1.5 hours at 110v. I will upload the gel images to GH tomorrow to get feedback on moving forward with the samples.