November 26, 2019 - Oyster Species Determination PCR and Gel Analysis & O. angasi Project

Oyster Species Determination - PCR and Gel Analysis

After looking at the results from the gel used to help determine the oyster species collected from the oyster distribution company (see last post here), Sam had discovered that the results from his gel were not was expected, since all of the samples were lined up in the same row (at around 500bps). After rerunning the PCR and gels with no multiplex, Sam had deduced that the wrong cytochrome oxidase primers were used, which is why there were no bands on the C. gigas- and C. sikamea-specific primers (see Sam’s post here).

So, Sam has ordered the correct primers to use, and we will begin the PCR process again tomorrow.

O. angasi Project

After talking with Dr. Roberts and Laura about potential research projects for next quarter, I have decided to help Laura analyze some samples of O.angasi that she acquired and tested during her time in Australia. In order to fully prepare for the project ahead of me, Laura has provided me with some information about the O. angasi samples and the experiments done.

Summary of the experiments done

  1. Heat Stress Experiment (Laura’s notebook entry here)
    • Heat shock trial was conducted on 2 groups of oysters and samples were collected during pre-stress, 6hr, and 24 hr markers of the experiment
    • Results: the conditions were shown to be lethal, since oysters began to die at ~48hrs after the shock trial, which is especially surprising since a previous experiment done on three other groups of oysters saw that the temperature was sub-lethal.
  2. Optimal Gonad Condition Experiment (Laura’s notebook entry here)
    • 3 temperature conditions tested (18C (control), 21C, and 24C) and food is constant to see the optimal conditions needed for gonad condition
    • Daily cleanings are done since previous experiments have shown that other species of oyster spawn more readily after cleaning
    • Results: gill, mantle, and gonad samples were collected and, overall, the oysters looked like they were in very good condition. Larvae were also collected during the experiment, mostly from the 24C treatment group.

Next Steps

  1. Repeat PCR and gel electrophoresis with new CO primers
Written on November 26, 2019