GSM4654589: sibetamcat_pos_24h_1; Homo sapiens; RNA-Seq
Cells were collected for RNA extraction using the RNeasy Mini Kit (Qiagen, 74104) according to the manufacturer's guidelines. RNA samples were quantified using RiboGreen (Invitrogen) on the FLUOstar OPTIMA plate reader (BMG Labtech) and RNA size profile and integrity were analysed on the 2200 or 4200 TapeStation (Agilent, RNA ScreenTape). RNA integrity number estimates for all samples were between 9.0 and 10.0. Poly(A) transcript enrichment and strand specific library preparation were performed using TruSeq Stranded mRNA kit (Illumina) following manufacturer's instructions. Libraries were amplified (15 cycles) on a Tetrad (Bio-Rad) using in-house unique dual indexing primers (Lamble et al., 2013). Individual libraries were normalized using Qubit and size profile was analysed on the 2200 or 4200 TapeStation. Individual libraries were pooled together and diluted to ~10 nM for storage. Each library aliquot was denatured and further diluted prior to loading on the sequencer.