GSM1406443: Gm6871-HA ChIP-seq in  mES cells; Mus musculus; ChIP-Seq
ES cell samples (ES3 cells overexpressing an HA-tagged Gm6871 expression vector) were washed 2x (in PBS + 2% FCS), counted to normalize by cell number, cross-linked (ten minutes rotation in 1% formaldehyde), quenched with glycine (at 125mM on ice), washed 3x (PBS) and pelleted at 10e7 cells per ependorf. Pellets were lysed, resuspended in 1ml sonication buffer on ice (10mM Tris pH 8, 200mM NaCl, 1mM EDTA, 0.5mM EGTA, 0.1% NaDOC, 0.25% NLS and protease inhibitors), transferred to glass 12x12mm tubes (Covaris: 520081) and sonicated (Covaris settings: 5% duty cycle, PIP 140, 200cyles/ burst, 20 minutes). Sonication was then assessed by reverse cross-linking overnight in the presence of proteinase K and RNase, followed by DNA extraction and quantification on a Bioanalyzer (Agilent 2100 machine). Libraries were prepared according to Illumina's instructions