Full day of sampling! THANK YOU to Grace for materials prep, Megan Hintz, Olivia & Lindsay Alma for helping Yaamini and me with the sampling, and to Steven for the oversight and OA system prep.

Here’s a nice example of an Oly, and the tissues we sampled:

The 4 populations have been in 2 temperature treatments (6 deg, ~10deg) for two months, and are slotted to go into OA treatments Feb 8th, so Feb 4th was a big sampling day.

Sampling scheme: all sampled animals were weighed (g), measured (mm), shucked, and tissue was extracted sparately:

• Mantle (M), ctenidia (C) & adductor (A) were sampled into 2mL centrifuge snap-top tubes, immediately flash-frozen in an ethanol/dry ice bath, and kept on dry ice until transported to the -80 in Rick’s lab.
• The remaining visceral mass sampled for gonad was put into histology cassettes, ~4 per cassette. Viscerall mass was placed in cassettes with right valve side facing up. I noted which specimens looked ripe. Cassettes were labeled using both histology pen and notches (see labeleing map), and put into the fixative within ~45 minutes of sampling.
• Any residual tissue (gill, mantle or adductor) was sampled for DNA by placing into tubes with 1mL ethanol, and stored at room temperature.

Number of each population sampled; see data sheet for details.

• Heare-2NF-6: 15
• Heare-2NF-10: 15
• Heare-2SN-6: 15
• Heare-2SN-10: 15
• Heare-2HL-6: 9
• Heare-2HL-10: 9

*None of Katherine’s were sampled today, due to histology fixative limitations.

Map of samples in histology cassettes:

An example of the varying ripeness within a population:

Inventory of supplies needed for sampling:

• scale
• calipers
• Sharp shucker - need to purchase/bring
• razor blade
• fine-nosed scissors
• Fine-nosed tweezers - need to purchase/bring
• Medium weight boats for dissections
• All-weather paper
• Bleach for sterilizing
• pre-labeled tubes, 4 per sample: Mantle, Ctenidia, Adductor, DNA tissue with 1ml ethanol
• pre-labeled casettes - need to figure out which labeling pen works!
• Ethanol
• Dry ice - enough for 2 coolers (for smooth organization & transfer of samples to -80)
• Glass container for dry ice bath
• Tweezers and panty hose for snap freezing in dry ice bath
• Tube racks
• Data sheets