GSM5195830: GAL117_025_02; Caenorhabditis elegans; RNA-Seq
Young adult hermaphrodites were picked using tweezers and washed in 0.5 mL sterile demineralized water before adding 200 μL trizol (Invitrogen 15596018). Samples were stored at -80 °C for a limited time. mRNA extraction, barcoding, reverse transcription, in vitro transcription and Illumina sequencing library preparation were performed according to the robotized version of the CEL-seq2 protocol (Hashimshony T. et al., 2016) using the SuperScript II Double-Stranded cDNA synthesis kit (Thermofisher), Agencourt AMPure XP beads (Beckman Coulter) and randomhexRt for converting aRNA to cDNA using random priming. The libraries were sequenced paired-end at 50 bp read length on an Illumina HiSeq 2500.