title: Sample 2
organism: Homo sapiens
description: Protocols: Nascent RNA was captured using 45min pulse according to the manual to Click-iT® Nascent RNA Capture Kit. 1 µg of DNAse-treated RNA was treated with RiboZero Gold (Human/Mouse/Rat) kit to remove rRNAs, followed by RNA cleanup using the RNeasy Minelute RNA cleanup kit. Sequencing libraries were prepared using TruSeq Stranded total RNA LT kit according to manufacturer's instructions using 11 cycles of PCR followed by purification with Agencourt Ampure XP beads. Libraries were evaluated on an Agilent 2100 Bioanalyzer using the DNA 1000 kit and DNA concentration was measured using a Qubit dsDNA HS Assay Kit. Samples were sequenced using a HiSeq2500 instrument to generate 50-nt single-end reads, sequencing depth was 20-40 Mio reads per library. H9 human ESCs (H9 line) were cultured in mTeSR1 medium on Matrigel-coated plates. Colonies were passaged for maintanence by Gentle Cell Dissociation Reagent Human ESCs were not treated and maintained in pluripotency using mTesr medium under same conditions and in parallel as samples prepared for total RNAseq. RNA-seq
attributes.tag: ENA first public
attributes.value: 2019-02-25
attributes.tag: ENA last update
attributes.value: 2016-10-03
attributes.tag: External Id
attributes.value: SAMEA4476484
attributes.tag: INSDC center alias
attributes.value: Institute of Molecular Life Sciences and Swiss Institute of Bioinformatics
attributes.tag: INSDC center name
attributes.value: Institute of Molecular Life Sciences and Swiss Institute of Bioinformatics
attributes.tag: INSDC first public
attributes.value: 2019-02-25T17:03:11Z
attributes.tag: INSDC last update
attributes.value: 2016-10-03T16:41:35Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-5115:Sample 2
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell type
attributes.value: undifferentiated human embryonic stem cell
attributes.tag: common name
attributes.value: human
attributes.tag: replicate
attributes.value: 2
attributes.tag: sample name
attributes.value: E-MTAB-5115:Sample 2