NuRD-mediated gene repression in mouse embryonic stem cells
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
No description
Antigen
NA
Cell type
Cell type Class
Pluripotent stem cell
Cell type
ES cells
NA
NA
Attributes by original data submitter
Sample
ENA-FIRST-PUBLIC
2012-03-13T17:00:57Z
ENA-LAST-UPDATE
2018-03-08T15:39:13Z
External Id
SAMEA1095979
INSDC center name
EMBL
INSDC first public
2012-03-13T17:00:57Z
INSDC last update
2018-03-08T15:39:13Z
INSDC status
public
Submitter Id
E-MTAB-888:Mbd3_WT2
broker name
ArrayExpress
cell type
embryonic stem cell
common name
house mouse
genotype
wild_type
sample name
E-MTAB-888:Mbd3_WT2
scientific_name
Mus musculus
Sequenced DNA Library
library_name
Mbd3_WT2_H3K27me3_ChIP
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
ES cells were grown in GMEM supplemented with 10% fetal bovine serum, 1 mM sodium pyruvate, 2 mM L-glutamine, 0.1 mM non-essential amino acids and 10 ng/ml murine LIF. Eed-/- ES cells were cultured on mitotically inactivated primary mouse embryonic fibroblasts which were depleted from ES cells upon harvest. TSA (Sigma) was added to normal ES cell media to a final concentration of 0.1 mM for 2 h prior to harvesting. Cells were fixed using 1% formaldehyde for 10min at room temperature and fixation quenched with 150 mM glycine. For Mi2beta ChIP-seq, formaldehyde fixation was preceded by 45 min incubation in 2 mM disuccinimidyl glutarate (Sigma). Chromatin was sheared by sonication to an average fragment size between 200 and 300 bp using a Bioruptor sonication instrument (Diagenode).