title: GSM1474187: WTTh1_p300; Mus musculus; ChIP-Seq
library_construction_protocol: Cells were fixed with 1% formaldehyde for 15 min and quenched with 0.125 M glycine. Chromatin was isolated by the addition of lysis buffer, followed by disruption with a Dounce homogenizer. Lysates were sonicated and the DNA sheared to an average length of 300-500 bp. Genomic DNA (Input) was prepared by treating aliquots of chromatin with RNase, proteinase K and heat for de-crosslinking, followed by ethanol precipitation. An aliquot of chromatin (15 ug for H3K4me3 and 25 ug for H3K4me1; 30 ug for all others) was precleared with protein A agarose beads (Invitrogen). Genomic DNA regions of interest were isolated using the antibodies listed. Complexes were washed, eluted from the beads with SDS buffer, and subjected to RNase and proteinase K treatment. Crosslinks were reversed by incubation overnight at 65 C, and ChIP DNA was purified by phenol-chloroform extraction and ethanol precipitation. Illumina sequencing libraries were prepared from the ChIP and Input DNAs using the Apollo 324 system (WaferGen). After a final PCR amplification step, the resulting DNA libraries were quantified and sequenced on HiSeq 2500.