Sample 72
Mus musculus
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).
ENA-FIRST-PUBLIC
2017-06-01T17:02:08Z
ENA-LAST-UPDATE
2017-04-19T13:59:31Z
External Id
SAMEA103983592
INSDC center name
EMBL-EBI
INSDC first public
2017-06-01T17:02:08Z
INSDC last update
2017-04-19T13:59:31Z
INSDC status
public
Submitter Id
E-MTAB-5661:Sample 72
broker name
ArrayExpress
cell line
OS25
cell type
embryonic stem cell
common name
house mouse
developmental stage
embryo
genotype
wild type genotype
growth condition
serum + LIF under Oct4 selection
phenotype
wild type
sample name
E-MTAB-5661:Sample 72
scientific_name
Mus musculus
strain
129/Ola