Sample Detail

TitleE-MTAB-2281:OTX2_8hr
DescriptionProtocols: Rex1GFPd2 mouse ES cells were maintained in NDiff N2B27 media in the presence of CHIR99021 and PD0325901 as described in Yang et al., 2012. For time series of differentiation, the media containing inhibitor was removed and replaced with NDiff N2B27 media as time lindicated. ES cells were cross-linked as described in Lee et al., 2006. Yang SH, Kalkan T, Morrisroe C, Smith A, Sharrocks AD. (2012) PLoS Genet. 2012;8(12):e1003112. A genome-wide RNAi screen reveals MAP kinase phosphatases as key ERK pathway regulators during embryonic stem cell differentiation. Lee TI, Johnstone SE, Young RA. (2006) Nat Protoc. 1(2):729-48. Chromatin immunoprecipitation and microarray-based analysis of protein location. ChIP-seq was performed as described in Lee et al., 2006, with the following modifications. The crosslinked material was sheared by water bath sonication using bio-rupter for 16 mins, at high level, 30 secs on/ 30 secs off, 4 ⁰C. The size of sheared DNA fragments was between 100 bp to 500 bp, with median size around 300 bp. Immunoprecipitation was performed by incubating 1.5×107cells with 2.5 µg of Otx2 antibodies (ProteinTech Group, Inc; 13497-1-AP) cross-linked to dynabeads protein A (Invitrogen), overnight at 4 ⁰C. Lee TI, Johnstone SE, Young RA. (2006) Nat Protoc. 1(2):729-48. Chromatin immunoprecipitation and microarray-based analysis of protein location.

Organism Info

Taxon ID10090
Common Name
Scientific NameMus musculus
Anonymized Name
Individual Name