alias: GSM2888688
title: GSM2888688: λH1-hESC derived_β-like cells; Homo sapiens; RNA-Seq
library_construction_protocol: RNA harvested from ~150,000 cells using Trizol reagent. (RNA processing, cDNA library construction and NGS sequencing were all performed at Ocean Ridge biosciences FL. USA) Total RNA was isolated from the tissue samples using the TRI Reagent® (Molecular Research Center; Part #: TR118) method. Following isolation, RNA was pooled from 3 human samples, quantified by O.D. measurement, and then digested with RNase free DNase I (Epicentre; Part # D9905K) and re-purified using RNeasy MinElute columns (Qiagen; Part #: 74204). The newly digested RNA samples were then quantified by O.D. measurement and checked for quality by chip-based capillary electrophoresis using an Agilent 2100 Bioanalyzer. Amplified cDNA libraries suitable for sequencing were prepared from 8 nanograms (ng) of DNA-free total RNA using the Clontech SMARTer Stranded Total RNA-Seq Kit v2 – Pico Input Mammalian (Clontech/Takara; Part # 634411). The quality and size distribution of the amplified libraries were determined by chip-based capillary electrophoresis (Agilent 2100 Bioanalyzer; Agilent Technologies). Libraries were quantified using the KAPA Library Quantification Kit (Kapa Biosystems, Boston, MA). The libraries were pooled at equimolar concentrations and diluted prior to loading onto the flow cell of the Illumina cBot cluster station. The libraries were extended and bridge amplified to create sequence clusters using the Illumina HiSeq PE Cluster Kit v4 and sequenced on an Illumina HiSeq Flow Cell v4 with 50 nt paired-end reads plus dual index reads using the Illumina HiSeq SBS Kit v4.
identifiers.namespace: GEO
identifiers.id: GSM2888688