characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2017-03-29T17:02:08Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2017-03-29T10:01:51Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA103946736
characteristics.characteristic: External Id
characteristics.text: Wellcome Trust Sanger Institute
characteristics.characteristic: INSDC center name
characteristics.text: 2017-03-29T17:02:08Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-03-29T10:01:51Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS1635904
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-5484:19287_7#12
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. A 96-well plate containing lysis buffer (0.2% Triton, 1:20 RNAse inbhibitor, 10mM Oligo-dT30VN, 10mM dNTPs) was prepared. Spike-ins (ERCCs and SIRVs; final 1:1000,000 dilution) are subjected to different freeze-thaw cycles (seven conditions) and 1µl of spike-in mix per condition was added to each well column-wise. Single mES cells were FACS sorted (BD Influx; BD Biosciences) to each well (along with bulk controls). Subsequent steps (Lysis, RT, PCR amplification and DNA purification) was performed as described in Smart-Seq2 publication (Picelli et al Nature Protocol 2014). The single cell cDNA across the 96-well plate was quantified, diluted 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: 19287_7#12
characteristics.characteristic: title