GSM8842321: T47DS_WT_R5020_4h_rep2; Homo sapiens; RNA-Seq
RNA isolation was performed using the RNeasy mini kit (Qiagen, #74104), including on-column DNAse digestion (Qiagen, RNase-free DNase set #79254) according to manufacturer's instructions. 1μg of RNA was used as starting material for poly-A enrichment using the NEBNext Poly(A) mRNA Magnetic Isolation Module (New England BioLabs). Next, RNA-Seq libraries were generated according to the manufacturers' protocols using the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina and NEBNext Multiplex Oligos for Illumina (Unique Dual Index Primer Pairs) (New England BioLabs). Distribution of the size of the libraries with indexed adapters was assessed using a 2200 TapeStation System with Agilent D1000 ScreenTapes (Agilent Technologies). The QuantStudio 3 Real-Time PCR System (Thermo Fisher Scientific) was used for library quantification using the NEBNext Library Quant Kit for Illumina (New England BioLabs) according to manufacturer's instructions.