GSM5088109: HepG2_Nelf_20uM_S13_rep1; Homo sapiens; RNA-Seq
Total RNA from four biological replicates per condition was extracted using the RNeasy Mini Kit (Qiagen) according to manufacturer's protocol for cultured cells, including on-column DNase I digestion. RNA yield and purity were measured on a NanoDrop 1000 Spectrophotometer (Thermo Scientific) to verify an A260/A280 nm ratio >1.8 and A260/A230 nm ratio ~2. Libraries were prepared from 250 ng of total RNA per sample using the Lexogen QuantSeq 3′ mRNA-Seq Library Prep Kit for Illumina (FWD) according to the manufacturer's instructions. Libraries were quantified with the Qubit dsDNA HS assay kit (Life Technologies) and combined in equimolar ratio. The pool of libraries was sequenced at the Estonian Genome Center Core Facility using the Illumina NextSeq500 platform with 80 bp single-end reads.