characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2017-06-01T17:02:08Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2017-04-19T13:59:31Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA103983588
characteristics.characteristic: External Id
characteristics.text: EMBL-EBI
characteristics.characteristic: INSDC center name
characteristics.text: 2017-06-01T17:02:08Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-04-19T13:59:31Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS1672756
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-5661:Sample 69
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: OS25
characteristics.characteristic: cell line
characteristics.text: embryonic stem cell
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/CL_0002322
characteristics.characteristic: cell type
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: Ring1A/B double knockout cells with constitutive Ring1A knockout and tamoxifen-inducible conditional Ring1B knockout were cultured on mitomycin-inactivated feeders in DMEM (lacking pyruvate; Gibco), supplemented with 10% batch-tested FCS (Labtech), 50mM ß-mercaptoethanol, L-glutamine (Gibco), Sodium Pyruvate (Gibco), Non-essential amino acids (Gibco), Penicillin/Streptomycin (Gibco) supplemented with 1000U per ml LIF (Milipore). These cell lines have been tested and were found to have no mycoplasma contamination. Feeders in Ring1AB dKO mES cells (untreated and tamoxifen-treated) are depleted using Feeder removal MicroBeads (Miltenyi Biotec). To induce Ring1b knockout, cells are cultured in media containing 800nM 4-hydroxytamoxifen (Sigma) for 48 hours and confirmed using genomic DNA isolation and PCR across Cre-excised region. OS25 cells were captured by loading the cell suspension on 10-17 micron C1 Single-Cell Auto Prep IFC, Fluidigm (without ERCC spike-ins). Ring1A knockout (untreated) and Ring1A/B double knockout cells are processed on C1 Single Cell Auto Prep System (Fluidigm; 100-7000 and 100-6209) using medium sized C1 mRNA-Seq chips (10-17 μm; 100-5670) with ERCC spike-ins (Ambion; AM1780) following the manufacturers protocol (100-5950 B1) requiring SMARTer kit for Illumina Sequencing (Clonetech; 634936). Single cell libraries were prepared using Illumina Nextera XT DNA sample preparation kit (Illumina; FC-131-1096) after cleanup and pooling using AMPure XP beads (Agencourt Biosciences; A63880).
characteristics.characteristic: description
characteristics.text: embryo
characteristics.characteristic: developmental stage
characteristics.text: wild type genotype
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0005168
characteristics.characteristic: genotype
characteristics.text: serum + LIF under Oct4 selection
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: wild type
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0005168
characteristics.characteristic: phenotype
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: 129/Ola
characteristics.characteristic: strain
characteristics.text: Sample 69
characteristics.characteristic: title