I sampled n = 20 newly hatched larvae from each experimental replicate on the day of peak hatch (March 4 for heatwave treatments and March 6 for ambient temp). Larvae from each replicate were preserved together in a 3 ml Eppendorf vial with RNALater and stored at -20C.

On June 17 2020, I selected a single larva from each vial, imaged it for morphometrics, and moved it to a fresh vial with 1 ml RNALater for transport to UW’s Functional Genomics Core. As a pilot, we plan to extract bulk RNA from single larvae to determine if the samples were adequately preserved to move forward with RNA-seq. The sample manifest can be found here.

I found that the condition of the larvae varied from vial to vial. Some samples appear more degraded than others and obtaining quality pictures for digital morphometric measurements was difficult (i.e., larvae had bent and shriveled in the preservative).