GSM9901076: NC_2; Homo sapiens; RNA-Seq
Total RNA was extracted from NC (negative control shRNA) and shCHI3L1 knockdown SKOV-3 human ovarian cancer cell lines using TRIzol reagent (Invitrogen) following the manufacturer's instructions. RNA integrity was assessed using Agilent 2100 Bioanalyzer (RIN value ≥7.0 for all samples), and RNA concentration was quantified by Qubit fluorometer. Genomic DNA contamination was removed by DNase I digestion before subsequent library construction. mRNA was enriched from qualified total RNA using oligo (dT) magnetic beads. Fragmentation buffer was added to interrupt mRNA into short fragments. First-strand cDNA was synthesized using random hexamer primers, followed by second-strand cDNA synthesis. After end repair, A-tailing and adapter ligation, fragments with suitable size were purified, PCR amplified to construct the final cDNA library. Paired-end sequencing was performed on Illumina sequencing platform.