GSM8322335: 2-786O-NEGsiRNA-RNAseq, rep2; Homo sapiens; RNA-Seq
Cells were lysed in TRIzol (15596018, Thermo Fisher Scientific) and snap frozen. RNA was then isolated using chloroform extraction, followed by QIAGEN Rneasy Mini Kit isolation (#74106), including DNA digestion with Dnase. Poly(A) + RNA was then isolated using double selection with poly-dT bead (E7490, NEB) RNA-seq libraries were prepared using a NEB-Next Ultra II Directional Lybrary Prep Kit for Illumina (E7760, NEB). Library sizes were determined on a Bioanalyzer, and concentrations determined using NEBNext Lib Quant Kit (E7630, NEB). Libraries were sequenced on an Illumina NextSeq 550, using paired-end sequencing of 42 bases per read.