alias: GSM3828594
title: GSM3828594: IPSC_Endoderm_8Day_01%DMSO_sample30; Homo sapiens; RNA-Seq
library_construction_protocol: Pelleted cells were snap frozen, then total RNA was extracted using the mirVana miRNA Isolation kit, with phenol (Ambion/ThermoFisher) according to manufacturer's protocol. Species specific rRNA-probes were hybridized to total RNA to remove nuclear-encoded and mitochondrial contaminating ribosomal RNA from the sample. The resulting ribo-depleted RNA was then fragmented. First-strand cDNA synthesis was performed using reverse transcriptase and random primers in the presence of Actinomycin D, followed by second-strand cDNA synthesis with DNA polymerase I and RNase H. Double-stranded cDNA was end-repaired and A-tailed for subsequent adaptor ligation. Indexed adaptors were ligated to the A-tailed cDNA. Enrichment by PCR was performed to generate the final cDNA sequencing library. Libraries were sequenced at a read depth of at least 30 million single-end 75 base pair reads on an Illumina NextSeq500 (San Diego, CA) following the manufacturer's protocols.
identifiers.namespace: GEO
identifiers.id: GSM3828594