GSM9698407: AO168O_2000_rep3; Caenorhabditis elegans; RNA-Seq
Approximately 1,000 worms per sample were washed thoroughly with M9 buffer, snap-frozen in liquid nitrogen, and stored at -80 °C until processing. Total RNA was extracted using TRIzol reagent (Invitrogen), and RNA integrity was assessed using an Agilent Bioanalyzer 2100 system. mRNA was enriched from total RNA using an Oligo(dT)-based workflow, followed by fragmentation, first-strand cDNA synthesis using random hexamer primers, second-strand cDNA synthesis, end repair, A-tailing, adaptor ligation, size selection, PCR enrichment, and purification. Libraries were initially quantified using Qubit 2.0, assessed for insert size using an Agilent 2100 Bioanalyzer, and further quantified by qRT-PCR before pooling and sequencing on an Illumina platform in paired-end 150-bp mode.