Today I performed a thermal stress experiment on Anthopleura elegantissima, with the goal of comparing the methylome and transcriptome before and after the stress.
Two specimens of A. elegantissima were brought to the lab from the sea tables at Shannon Point Marine Center. These individuals were originally collected from Lawrence Point, Orcas Island, WA, in late June 2014. They were brought to the Roberts Lab on 2/3/2015, and were kept in a small plastic container with about 1L of seawater, which was changed weekly. The container was held inside a well-lit refrigerator kept at 5C.
Today, after the specimens had spent two weeks under these conditions, I clipped a few tentacles from each of them before transferring them into beakers filled with seawater, which were then placed inside a water bath at 37C for one hour. After this thermal stress, I again clipped a few tentacles. Before and after tentacle samples were immediately placed in the -80C freezer. The samples were placed in a freezer box on the bottom shelf of the freezer.
The larger of the two anemones was labelled as “#1″, while the smaller one was labelled “#2″. After the heat shock, both anemones had detached from the watch glasses they had been attached to, indicating that they were clearly affected by the shock. The larger of the two anemones appeared slightly healthier after the shock; I suspect it is the one more likely to survive if there is any mortality.
I came in to find that both anemones had died. They were not heavily decomposed so either they died recently, or were kept in good shape by the cold water temperature.
I perfomed DNAzol isolation of DNA on the samples. Only two of the samples showed an obvious precipitate, but because of the generally low quantities I opted to pellet the DNA instead of spooling it. I left the samples in 95% ethanol and plan to quantitate the DNA next week.