S.mediterranea RNASeq
Animals of sexual S. mediterranea were homogenized in Trizol reagent (Invitrogen) on ice using a Miccra D-8 homogenizer (Miccra) and processed according to the Trizol reagent protocol. Following phase separation by chloroform addition, and isopropanol precipitation, the RNA pellet was dissolved in 350 Âľl RLT buffer and purified according to the RNeasy Mini Kit (Qiagen), including on-column DNase I digestion. The RNA was eluted in nuclease-free water (Ambion) and stored at -80 Â°C. The total RNA was quantified using a NanoDrop 1000 spectrophotometer and integrity was checked by capillary electrophoresis on a Bioanalyzer 2100 (Agilent) using the RNA 6000 Nano Kit and processed for stranded RNAseq library preparation and sequencing.