title: hESCs HERVH knockdown
organism: Homo sapiens
description: Protocols: The hESC line H1 (WA-01, passage 30) was cultured feeder-free in conditioned medium on Matrigel. The hESCs were passaged for expansion with 1 mg/ml collagenase IV (Gibco) every 5-7 days. For transfection, the hESCs were passaged with 0.25% trypsin (Invitrogen) for dissociation and transfected with 2 ug shRNA and 6 ul Mirus LT1 transfection reagent the following day. Trizol (Invitrogen) was used for extraction of total RNA from hESCs treated with control shRNA or HERVH shRNA for 5 days. RNA extracted from multiple samples was pooled. 4 ug total RNA was poly (A) enriched by incubating with oligo(dT) coated sepherose beads for two rounds (Ambion). The resultant enriched mRNA was fragmented, size selected and reverse transcribed according to the TruSeq RNA and DNA Sample Preparation Kits v2 (Illumina).
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2014-03-30T17:00:36Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-03-08T16:52:49Z
attributes.tag: External Id
attributes.value: SAMEA2248195
attributes.tag: INSDC center name
attributes.value: GIS
attributes.tag: INSDC first public
attributes.value: 2014-03-30T17:00:36Z
attributes.tag: INSDC last update
attributes.value: 2018-03-08T16:52:49Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-2072:hESCs HERVH knockdown
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell line
attributes.value: H1
attributes.tag: cell type
attributes.value: embryonic stem cell
attributes.tag: common name
attributes.value: human
attributes.tag: sample name
attributes.value: E-MTAB-2072:hESCs HERVH knockdown
attributes.tag: scientific_name
attributes.value: Homo sapiens
attributes.tag: sex
attributes.value: male