characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2017-03-28T17:01:34Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2017-03-28T16:26:04Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA103944242
characteristics.characteristic: External Id
characteristics.text: Wellcome Trust Sanger Institute
characteristics.characteristic: INSDC center name
characteristics.text: 2017-03-28T17:01:34Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-03-28T16:26:04Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS1633410
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-5483:19083-C1_7#55
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: ES-E14
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0007075
characteristics.characteristic: cell line
characteristics.text: mouse embryonic stem cell
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0004038
characteristics.characteristic: cell type
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: Wildtype E14 mESC were cultured under standard culture conditions in Knockout DMEM containing 15% Fetal Calf Serum, 1x Penicillin-Streptomycin-Glutamine, 1x NEAA 2-mercaptoethanol and 1000U Leukemia Inhibitory Factor. Trypisnised single mESC suspension is passed through 30micron filter to remove and clumps. Scripts with different method workflows were downloaded from Fluidigm Script Hub. Approx 3000 single mESC are loaded onto 5-10 micron Fluidigm C1 Single-Cell Auto Prep IFC and cell capture was performed as per manufacturers protocol. Two kinds of spike-ins (ERCC-#4456740 and SIRV-E2 #SKU025.03) were mixed (0.5ul 1:500 diluted ERCCs + 0.6ul 1:500 diluted SIRVs) and added to respective Lysis buffer. The subsequent steps are performed as per manufacturers protocol or described on Fluidigm Script hub. The cDNA from each single chamber within C1-IFC was harvested and diluted as per manufacturer protocol. Harvested cDNA from single cells was quantified and libraries were made using Nextera XT DNA sample preparation kit (Illumina) utilising 96 dual barcoded indices. Library pooling was performed post AMPure XP beads (Agencourt Biosciences) clean up and sequenced on HiSeq2500 (v4) using paired end sequencing (75 or 125bp) at the Wellcome Trust Sequencing facility following Wellcome Trust Sequencing guidelines.
characteristics.characteristic: description
characteristics.text: Serum + LIF condition
characteristics.characteristic: growth condition
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_10090
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: 129/ola
characteristics.characteristic: strain
characteristics.text: 19083-C1_7#55
characteristics.characteristic: title