Illumina HiSeq 2000 sequencing; Otx2 binding to Nanog promoter protects embryonal stem cells from corruption of naive pluripotency and is required at morula stage for inner-cell mass differentiation of the epiblast
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-CHECKLIST
ERC000011
ENA-FIRST-PUBLIC
2016-06-09T17:01:07Z
ENA-LAST-UPDATE
2018-03-09T03:28:22Z
External Id
SAMEA3522260
INSDC center name
Telethon Institute of Genetics and Medicine (Tigem)
INSDC first public
2016-06-09T17:01:07Z
INSDC last update
2018-03-09T03:28:22Z
INSDC status
public
Submitter Id
E-MTAB-3856:ESC_WT1
broker name
ArrayExpress
cell line
embryonic cell line
cell type
embryonic stem cell
common name
house mouse
genotype
wild type genotype
sample name
E-MTAB-3856:ESC_WT1
scientific_name
Mus musculus
Sequenced DNA Library
library_name
ESC_WT1
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
ChIP-assay were performed from 10e7 cells per experiment. Cells were crosslinked with 1% formaldehyde for 10 min at room temperature. Glycine at a final concentration of 0.125 M was added to quench the reaction. Cells were collected and lysed in SDS buffer and chromatin was then sonicated to an average size of 0,2-0,3 kb. After a 30 minute step of chromatin preclearing with the Protein A Agarose/Salmon Sperm DNA 50% gel slurry (Millipore), rabbit polyclonal antibody against Otx2 was added to the chromatin and incubated overnight at 4°C. The immunoprecipitated complex was subsequently incubated with the Protein A Agarose for 2-4 hours at 4°C. Agarose beads were then washed, chromatin was eluted and reverse-crosslinked overnight at 65°C. E14Tg2a wt and Obs3 mutant ESCs were routinely grown in GMEM medium plus 12% FBS (Hyclone) and LIF (Millipore). Proteinase K treatment and phenol/chloroform purification. Libraries were prepared according to Illumina standard protocols with internal barcodes.