GSM5548071: HGD-5 biol rep 4; Caenorhabditis elegans; RNA-Seq
Pelleted worms were lysed by beat beating in TRIzol reagent (Thermo Fisher, Waltham, MA). Total RNA was isolated using Nucleospin RNA (Macherey Nagel) following the manufacturer's protocol. Contaminant genomic DNA was removed from samples with TURBO DNase (Thermo Fisher) following the manufacturer's protocol. mRNA quality was assessed by NanoDrop 1000 v3.8 (Thermo Scientific), and 100 ng/µl of the samples with RNA integrity number above 8.0 were proceeded with library preparation. The total RNA quality and RIN value was confirmed with a Tapestation from Agilent and Agilent RNA ScreenTape. Library preparation was done using a commercially available kit, Illumina's TruSeq® Stranded mRNA Library Prep, following the manufacturer's protocol. The different libraries were then pooled together and QC using Agilent high sensitivity DNA kit and KAPA quantification.