RNAseq of Elysia crispata: egg
Total RNA was isolated from 100 mg of pulverized flash-frozen E. crispata tissue using Trizol reagent (ThermoFisher Scientific, Waltham, MA) according to the manufacturers protocol. Extracted RNA samples were digested with DNaseI (New England Biolabs, Ipswich, MA) and column purified using an RNA concentrator and cleanup system (Zymo Research, Irvine, CA). Purified samples were measured for RNA integrity (RIN) scores at the Purdue Genomics Core facility. Samples with RIN score > 7 were selected for library prep. Three biological replicates were sent to Novogene Corporation Inc. (Sacramento, CA) for library construction (NEBNext Ultra RNA Library Prep Kit, New England Biolabs) from 1 g total RNA and Illumina sequencing.