GSM3613316: Control lateG1-2; Homo sapiens; RNA-Seq
For bulk-population RNA-seq, RNA was extracted from cell cycle sorted control and DMSO-treated hPSCs.  Integrity of extracted RNA was assayed by on-chip electrophoresis (Agilent Bioanalyzer) and only samples with a high RNA integrity (RIN) value were used for RNA-seq.  Purified total RNA was reverse-transcribed into cDNA using the Ovation RNA-seq System V2 (NuGEN) and cDNA was sheared using the Covaris S2 system (duty cycle 10%, intensity 5, cycle/burst 100, total time 5 min). Sheared cDNA was cleaned up using Agencourt AMPure XP beads (Beckman Coulter) and ligated to adaptors (Illumina) Sequencing libraries were constructed using the NEBNext Ultra DNA Library Prep Kit (New England Biolabs) using barcoded adaptors to enable multiplexing of libraries on the same sequencing lane. For each RNA-seq library, the effectiveness of adaptor ligation and effective library concentration was determined by Bioanalyzer before loading them in multiplexed fashion onto an Illumina HiSeq4000 (Stanford Functional Genomics Facility) to obtain ~150 bp paired-end reads.
GEO Accession
GSM3613316