GSM2123552: Nanog ChIP (haploid); Mus musculus; ChIP-Seq
ChIP-seq samples were prepared as described in Morey et al, 2015. Briefly, 5 million mESCs were removed from the dish using accutase and resuspended in 2i media. mESCs were then fixed in 1 % formaldehyde for 10 min before adding 0.0125 M glycine for 5 min to quench the reaction. Chromatin was then immunoprecipitated with 5 ug of antibody Libraries were prepared according to Illumina's instructions accompanying the DNA Sample Kit (Part# 0801-0303). Briefly, DNA was end-repaired using a combination of T4 DNA polymerase, E. coli DNA Pol I large fragment (Klenow polymerase) and T4 polynucleotide kinase. The blunt, phosphorylated ends were treated with Klenow fragment (32 to 52 exo minus) and dATP to yield a protruding 3- 'A' base for ligation of Illumina's adapters which have a single 'T' base overhang at the 3’ end. After adapter ligation DNA was PCR amplified with Illumina primers for 15 cycles and library fragments of ~250 bp (insert plus adaptor and PCR primer sequences) were band isolated from an agarose gel. The purified DNA was captured on an Illumina flow cell for cluster generation. Libraries were sequenced on the Genome Analyzer following the manufacturer's protocols.
GEO Accession
GSM2123552