GSM8290064
Worms were collected in TRIzol Reagent (Invitrogen), flash frozen in liquid nitrogen, and stored at -80°C prior to RNA isolation. Samples were lysed using metal beads and the Tissuelyser I system (Qiagen). RNA was isolated from lysate using Direct-zol RNA Miniprep kit (Zymo) and stored at -80°C prior to library preparation. Total RNA samples were evaluated for quality control using a NanoDrop One C Microvolume UV-Vis Spectrophotometer (ThermoFisher Scientific), with an A260/A280 > 1.9 and an A260/A230 > 2.2. Approximately 1 ug of RNA per sample was submitted to Genewiz (Azenta) for QC, library preparation, and total mRNA sequencing. All total RNA quality control, mRNA-sequencing library preparation, and Illumina sequencing was performed by Genewiz (Azenta). Samples were first verified for RNA integrity scores greater than 8.5 using an Agilent Tapestation 4200, and a DV200 score greater than 70.0. Illumina library preparation was performed using polyA selection for mRNA species. Approximately 20 million paired-end, 150 base pair reads were obtained per sample, with ≥ 80% of bases passing a Phred quality score ≥ Q30.