GSM9285129: HepaRG cells, siADARB1, replicate 3; Homo sapiens; RNA-Seq
Total RNA was extracted using TRI Reagent® (Zymo Research, CA, USA) and purified using the Direct-zol RNA Miniprep kit (Zymo Research, CA, USA For library preparation, ribosomal RNAs were removed by Illumina epidemiology rRNA Removal Kits, and the RNA-seq library was processed using NEBNext® Ultra™ II Directional RNA Library Prep Kit for Illumina (NEB, USA) following the manufacturer's recommendations. The library was barcoded, enriched, and purified using Agencourt AMPure beads (Beckman Coulter, catalog # A63881). The size and mass of the library were assessed using the Agilent TapeStation with a DNA5000 Screen Tape. Quantitative PCR was used to validate the library's functionality using the NEBNext Library Quantification kit for Illumina (NEB, E7630S). The libraries were pooled at equal molar concentrations and sequenced on an Illumina NovaSeq 6000 at the University of Florida ICBR NextGen Sequencing Core (RRID: SCR_019152). The pooled library was diluted to 0.8 nM and sequenced on one S4 flow cell lane (2 x 150 cycles) with 1% PhiX spike-in control.