GSM8380977: WIDR-SH1-1; Homo sapiens; RNA-Seq
Total RNA was extracted using the TRIzol reagent kit (Invitrogen, Carlsbad, CA, USA) according to the manufacturer's protocol. RNA quality was assessed on an Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA, USA) and checked using RNase-free agarose gel electrophoresis. After total RNA was extracted, eukaryotic mRNA was enriched using Oligo(dT) beads. The enriched mRNA was fragmented into short fragments using fragmentation buffer and reverse-transcribed into cDNA using the NEBNext Ultra RNA Library Prep Kit for Illumina (NEB#7530, New England Biolabs, Ipswich, MA, USA). The purified double-stranded cDNA fragments were end-repaired, A-tailed, and ligated to Illumina sequencing adapters. The ligation reaction was purified with AMPure XP Beads (1.0X) and PCR amplified. The resulting cDNA library was sequenced using the Illumina Novaseq 6000 (Repugene Technology, Hangzhou, China).