GSM9245732: Caco-2 cells, Nsp3mut, 20 h, biol rep 3; Homo sapiens; RNA-Seq
Total RNA was extracted using the E.Z.N.A. HP Total RNA Kit Strand-specific RNA-seq libraries were prepared with the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (New England Biolabs) following the manufacturer's instructions. Briefly, enriched RNAs were fragmented for 8 min at 94 °C, followed by first- and second-strand cDNA synthesis. Strand specificity was preserved by incorporation of dUTP during second-strand synthesis. cDNA fragments were adenylated at 3′ ends, ligated to indexed adapters, and enriched by limited-cycle PCR. Libraries were validated on an Agilent TapeStation, quantified by Qubit 3.0 Fluorometer and qPCR (KAPA Biosystems), multiplexed, and sequenced on an Illumina NovaSeq X Plus using a 2×150 bp paired-end configuration.