GSM5290475: QuantSeq_RNAseq_MM1S_NCOR1_sgRNA2_DMSO_6hours_replicate1; Homo sapiens; RNA-Seq
For 3'-mRNA-seq (QuantSeq), Cells were collected and washed once with ice-cold PBS prior to resuspension in TRIzol™ (ThermoFisher Scientific, 15596026). RNA was isolated using the Direct-zol RNA MiniPrep kit (Zymo Research, R2052) according to the manufacturers instructions. Sequencing libraries were prepared from 500ng RNA using the QuantSeq 3'-mRNA Seq Library Prep Kit for Ilumina (Lexogen, Vienna, Austria). Libraries were sequenced on the Illumina NextSeq 500 to obtain 75 b.p. single-end reads. For Total RNA-seq, Total RNA was immediately extracted using TRIzol reagent (ThermoFisher Scientific, 15596026) and isolated using the Direct-zol RNA MiniPrep kit (Zymo Research, R2052). Libraries were prepared with NEBNext Ultra II Directional RNA Library Prep Kit for Ilumina (NEB, E7760S) with ribosomal RNA depletion and sequenced on an Ilumina NextSeq 500 with 75 b.p. single-end reads.