GSM1681920: Donor 1 - H3K27ac; Homo sapiens; ChIP-Seq
In order to perform ChIP experiments the chromatin of day 11 cells was cross-linked for 8 min with 1% formaldehyde in PBS. Then the formaldehyde was quenched for 5 min with 125 mM of glycine. Nuclei were isolated and the chromatin was sheared with 30 cycles at high intensity (30 sec off and 30 sec on) using a sonicator (BioruptorTM Next Gene, Diagenode). For each immunoprecipitation 5 μg of sheared chromatin was used. Pre-cleared chromatin was incubated overnight with 5 μg of an antibody against the active enhancer mark H3K27ac (Abcam; ab4729). Following day the antibodies were captured with 25 μl of protein A magnetic beads (Millipore)s for 2 hours on a rotating wheel at 4°C. The DNA was purified with a QIAquick PCR clean-up kit. ChIP sequencing was performed at the GeneCore facility of EMBL Heidelberg (http://www.genecore.embl.de/) according to the Illumina protocol.
GEO Accession
GSM1681920