characteristics.text: 2013-08-30T17:00:54Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2018-03-08T16:29:47Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA2078456
characteristics.characteristic: External Id
characteristics.text: GIS
characteristics.characteristic: INSDC center name
characteristics.text: 2013-08-30T17:00:54Z
characteristics.characteristic: INSDC first public
characteristics.text: 2018-03-08T16:29:47Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS252028
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-1687:hESCs
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: H1
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0003042
characteristics.characteristic: cell line
characteristics.text: embryonic stem cell
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/CL_0002322
characteristics.characteristic: cell type
characteristics.text: human
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_9606
characteristics.characteristic: common name
characteristics.text: 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.
characteristics.characteristic: description
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: Homo sapiens
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_9606
characteristics.characteristic: organism
characteristics.text: Homo sapiens
characteristics.characteristic: scientific_name
characteristics.text: male
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/PATO_0000384
characteristics.characteristic: sex
characteristics.text: hESCs
characteristics.characteristic: title