GSM2232860: H3K9me3 Naive lane 2; Homo sapiens; ChIP-Seq
Pellets were lysed, resuspended in 1mL of LB1 on ice for 10min (50 mM HEPES-KOH pH 7.4, 140 mM NaCl, 1 mM EDTA, 0.5 mM EGTA, 10% Glycerol, 0.5% NP40, 0.25% Tx100, protease inhibitors), then after centrifugation resuspend in LB2 on ice for 10min (10 mM Tris pH 8.0, 200 mM NaCl, 1 mM EDTA, 0.5 mM EGTA and protease inhibitors). After centrifugation, resuspend in LB3 (10 mM Tris pH 8.0, 200 mM NaCl, 1 mM EDTA, 0.5 mM EGTA, 0.1% NaDOC, 0.1% SDS and protease inhibitors) and sonicated (Covaris settings: 5% duty, 200 cycle, 140 PIP, 60 min), yielding genomic DNA fragments with a bulk size of 100-300 bp Libraries of immunoprecipitated chromatin and total input control from ChIP were performed with single-end adaptors as previously described (Rowe et al., 2013b). Sequencing was performed on an Illumina HiSeq 2500 (Illumina), with each library sequenced in 100-bp reads run.