GSM3391427: hsb-1(-) rep3; Caenorhabditis elegans; RNA-Seq
TRIzol reagent (Thermo Fisher) was used to extract total RNA from ~1,000 age-synchronized day 1 adult worms (three biological replicates per genotype). DNase-treatment was performed using TURBO DNA-free kit (Themo Fisher). mRNA was isolated from the DNase-treated total RNA using oligo(dT) and cDNA was synthesized by reverse transcription using TruSeq RNA Library Prep Kit v2 (Illumina). The samples were end-repaired, poly(A)-tailed, ligated to barcoded oligo adapters (Illumina) and PCR-amplified using Apollo 324 library preparation system (Wafergen Bio-systems) to prepare non-stranded poly(A)-based cDNA libraries with 120 bp insert size.