title: GSM8196109: RNAseq of total lysate NAC_KD replicate 1; Caenorhabditis elegans; RNA-Seq
library_construction_protocol: Frozen worm pellets were lysed using cryo-mill with lysis buffer without detergent. For total RNA, detergent was added to thawed lysate, spun at 21,000g for 15 minutes to clear the lysate, and RNA was extracted using TRIzol reagent following manufacturer's instructions. Soluble fraction was separated by thawing lysate without detergent, spinning at 21,000g for 15 minutes, and RNA was extracted using TRIzol reagent. Lysis buffer with detergent was added to the remaining pellet from isolating soluble fraction, incubted for 10 min, spun at 21,000g for 15 minutes to isolate detergent-extracted membrane fraction, and RNA was extracted using TRIzol reagent. Library was prepared using TruSeq Stranded Total RNA Prep kit (Illumina) following protocol provided by manufacturer. rRNA was depleted using Ribo-Zero rRNA removal kit (Illumina).