organism: Homo sapiens
description: Protocols: The hESC line H1 (WA-01, passage 30) was cultured feeder-free in conditioned medium on Matrigel (BD). Conditioned medium contains 20% Knock-out serum replacement (Gibco), 1 mM L-glutamine (Gibco), 1% non-essential amino acids (Gibco) and 0.1 mM 2-mercaptoethanol (Gibco), 4 ng/ml human basic fibroblast growth factor (bFGF) (Invitrogen), and medium was supplemented with additional bFGF (8ng/ml) before use. 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 1.5 ug shRNA and 4.5 ulMirus LT1 transfection reagents the following day. Cells were harvested for mRNA 3 days after transfection of SON shRNA1. Trizol (Invitrogen) was used for extraction of total RNA from hESCs treated with control shRNA or SON shRNA for 3 days. 4 ug RNA extracted from biological duplicate samples 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 IlluminaTruSeq RNA Sample Preparation Guide.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2013-08-30T17:00:54Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-03-08T16:29:47Z
attributes.tag: External Id
attributes.value: SAMEA2078456
attributes.tag: INSDC center name
attributes.value: GIS
attributes.tag: INSDC first public
attributes.value: 2013-08-30T17:00:54Z
attributes.tag: INSDC last update
attributes.value: 2018-03-08T16:29:47Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-1687:hESCs
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-1687:hESCs
attributes.tag: scientific_name
attributes.value: Homo sapiens
attributes.tag: sex
attributes.value: male