GSM9568332: Pooled primary human hepatocytes, replicate 2; Homo sapiens; RNA-Seq
Total RNA was isolated using the QIAwave RNA Kit (50) (74534, Qiagen) according to the manufacturer's instructions. Adherent monolayers were washed with PBS and lysed directly in RLT buffer supplemented with β-mercaptoethanol (1% v/v). For pooled PHH, cryovials were thawed (37 °C, 2 min), diluted in nine volumes of ice-cold PBS, pelleted (100 × g, 5 min, 4 °C), supernatant removed, and pellets lysed in RLT buffer. RNA concentration and purity (A260/280 and A260/230) were assessed using a NanoDrop 2000 UV-Vis spectrophotometer. RNA-seq libraries were prepared and sequenced by Eurofins Genomics (INVIEW™ Transcriptome Discover service). mRNA was enriched using bead-based poly(A) selection, followed by library preparation (Eurofins standard pipeline). Libraries were sequenced on an Illumina NovaSeq platform as paired-end 2 × 150 bp reads (NovaSeq PE150) with a target yield of 30 million read pairs per sample (±3%).