Alternative splicing of circadian genes in intertidal copepods

This study aimed at characterizing splice variants associated with circadian rhythms in intertidal copepods.The experiment used copepods from San Diego, California, and Strawberry Hill, Oregon that had been acclimated to laboratory conditions for at least three generations.Copepods were cultured in an incubator at 20C in one of two different light regimes: 8:16 or 12:12: Light:Dark cycle. For each treatment (population x light regime), 25-day old adult male individuals were sampled in pools of ~20 individuals five hours after the light turned on. Five replicates were collected from each treatment (total 20 samples).Total RNA was isolated using Trizol reagent, and sequencing library was prepared with the NEBNext Ultra II Directional RNA kit, using 300-500 ng of total RNA as input.