GSM6742731: SI-SCR_B; Homo sapiens; RNA-Seq
Total RNA was extracted from iPSC-derived NPC lines using TRIZOL reagent (Invitrogen) following the manufacturer's instruction. DNA contamination was removed by treating the samples with DNase I (Invitrogen) and RNase inhibitor SUPERase (Invitrogen) for 30' at 37°C followed by purification with RNase Mini Kit (Qiagen). RNA quality was evaluated by agarose gel electrophoretic run and Agilent 2100 Bioanalyzer. RNA-seq libraries from hnRNPL si-RNA treated samples (si-hnRNPL-C and si-scrambled control (si-SCR) were obtained from total RNA isolated using TRIZOL (Invitrogen) and treated with DNase (AMBION)). RNA quality was assessed by Agilent bio-analyzer (RIN from 7.8 to 9.2). Selected mRNA on magnetic oligo(dT) beads were retro-transcribed using random hexamer. A second strand was generated by dUTP method. Standard Illumina paired-end (100bp) library preparation was performed with barcoded adaptors to obtain ~50-60 million reads per sample.