characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2014-12-30T17:01:23Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2018-03-08T19:44:06Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA2699628
characteristics.characteristic: External Id
characteristics.text: EMBL-EBI, Wellcome Trust Genome Campus, Cambridge
characteristics.characteristic: INSDC center name
characteristics.text: 2014-12-30T17:01:23Z
characteristics.characteristic: INSDC first public
characteristics.text: 2018-03-08T19:44:06Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS527362
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-2805:S_cell87
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: S
characteristics.characteristic: cell cycle stage
characteristics.text: AB2.2
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0002812
characteristics.characteristic: cell line
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: mESC were grown in standard 2i media. The media composition was N2B27 basal media (NDiff 227, StemCells), 100 U/ml recombinant human leukemia inhibitory factor (Millipore), 1M PD0325901 (Stemgent), 3M CHIR99021 (Stemgent). For sorting of cell cycle fractions (G1, S and G2M), cells were trypisinized and stained with Hoechst 33342 (5_M) for 30min at 37C. Cells were sorted based on Hoechst staining for G1, S and G2M stages of cell cycle. For each cell cycle fraction, 3000 cells were loaded onto a 10-17 micron Fluidigm C1 Single-Cell Auto Prep IFC and cell capture was performed as per manufacturers protocol. Single cell capture, lysis, Reverse transcription and cDNA amplification are performed using the SMARTer cDNA synthesis kit (Clonetech) and Advantage2PCR kit (Clonetech) with 1:100 ratio of spike in (Ambion), within the IFC inside the C1-Autoprep system as per manufacturers protocol. Harvested cDNA from single cells after C1-Autoprep system run were used to make single cell libraries using Nextera XT DNA sample preparation kit (Illumina) utilising 96 dual barcoded indices. Library poolup was performed using AMPure XP beads (Agencourt Biosciences) and single cell libraries were sent for paired end sequencing at the Wellcome Trust Sequencing facility
characteristics.characteristic: description
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: fresh specimen
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/OBI_0000971
characteristics.characteristic: specimen with known storage state
characteristics.text: mESC
characteristics.characteristic: strain
characteristics.text: S_cell87
characteristics.characteristic: title