GSM9901875: Liver, EtOH-fed Ripk1-hepKO female mouse, S37; Mus musculus; RNA-Seq
Total RNA was extracted from mouse liver tissues using TRIzol reagent (Invitrogen) according to the manufacturer's instructions. RNA concentration and purity were evaluated using a NanoDrop 2000 spectrophotometer (Thermo Fisher Scientific) by measuring the A260/A280 and A260/A230 ratios. Before library construction, RNA integrity was assessed by agarose gel electrophoresis and RNA quantity was further assessed using Qubit fluorometric quantification. Polyadenylated mRNA was enriched from total RNA using Oligo(dT) magnetic beads and randomly fragmented into approximately 200-bp fragments. First-strand cDNA was synthesized using random primers. During second-strand cDNA synthesis, dTTP was replaced by dUTP to enable strand-specific library construction. Double-stranded cDNA was purified using AMPure XP beads, end-repaired, A-tailed, and ligated to sequencing adapters. The second cDNA strand was digested using USER enzyme, and the resulting strand-specific libraries were enriched by approximately 15 cycles of PCR. Libraries were quality assessed and quantified before paired-end 150-bp sequencing on an Illumina NovaSeq 6000 platform.