GSM1567362: H3K27me3 ChIP-seq; Mus musculus; ChIP-Seq
Freshly dissected mouse sciatic nerve was minced in 1% formaldehyde for 10min and then quenched for 10min with glycine to a final concentration of 0.125M. Samples were sequentially lysed in buffers LB1, LB2, and LB3 + 0.03% SDS (Schmidt et. al., 2009, PMCID:  PMC4052679).   DNA was fragmented to an average size of 0.5-2kb using 4x10min Bioruptor (Diagenode) cycles on the medium setting.  Each aliquot of sonicated chromatin (300ug) was incubated overnight at 4°C with 5ug of H3K27me3 antibody (Millipore 07-449, Lot#2064519).  A 10% aliquot was saved for input analysis.  An 80ul aliquot of Protein G Dynabead (Invitrogen) slurry was added to each ChIP sample for 4hrs at 4°C.  IPs were washed 5x in RIPA buffer than eluted at 65°C with reverse crosslinking buffer (50mM Tris, 10mM EDTA, 1% SDS).    ChIP DNA was purified by phenol chloroform extraction and resuspended in 10mM Tris pH 8.0. Libraries were prepared from  H3K27me3 ChIP sample, and corresponding input, using the Illumina TruSeq ChIP Sample Preparation Kit.