Prepping the remaining mussel samples for DNA extraction. See notes in post.

Method notes:

Link to steps I did for sampling: here

The samples were saved from the experiment in the same bags they were kept in for the experiment. They were stored at -80C.

I pulled the mussel bags out and let thaw to room temp for 1.5hours.

To sample the mussels, I opened the bag and layed it flat on the benchtop. I slowly pushed the mussel up towards the opening, making one side such that it didn’t scrape along the inside of the bag on the way up. I then took the shell swab sample (10s with a flocked cotton swab along the side of the shell that wasn’t scraped along the inside of the bag), and put the swab in a 1.5mL snap cap tube for storage in -80C.

To sample the tissue, I used sterilized tools (wiped with a kimwip; 10% bleach, 70% EtOH, flame) to pull out the mussel tissue into a 1.5mL snap cap tube. I used a poker tool from my dissection kit (sterilized between uses) to mash up the mushy tissue in the tube. I then used sterilized tweezers and scissors to put 150mg of tissue into a new 1.5mL snap cap tube (used the scale in FTR 213 to weigh to ~0.15 g –> actual measurements recorded in sample log). The remaining tissue was kept in the original 1.5mL snap cap tube.

I put the swabs, remaining mussel tissue samples, and the ~150mg tissue samples into the -80C.

I had to re-freeze samples M_EM_03; M_MSW_01; M_MSW_02 because the lighter ran out of juice yesterday to sterilize tools. I thawed them again today and finished the swabbing and tissue sampling from theses samples.

Next steps:

Extract DNA from:

• mussel shell swabs (I have the kit!)
• mussel tissue (150mg of tissue per sample)