GSM2419205: H3K27ac_LPS_1h; Mus musculus; ChIP-Seq
Approximately 10x10^6 of untreated or stimulated (1h and 4h with LPS) RAW cells were used for each experiment. Cells were crosslinked with 0.75% formaldehyde for 10 min at RT and chromatin was sonicated as described previously (Soldi et. Al, 2013). Each chromatin input was immunoprecipitated with 10 g of antibody. beads were washed in Buffer A (20 mM Tris-HCl pH 7.6, 2 mM EDTA, 0.1% SDS, 1% Triton-100, 150 mM NaCl), Buffer B (20 mM Tris-HCl pH 7.6, 2 mM EDTA, 0.1% SDS, 1% Triton-100, 300 mM NaCl) and once in TE containing 50 mM NaCl. DNA was eluted in TE containing 2% SDS and de-crosslinked overnight at 65°C. DNA was then purified by Qiaquick columns (QIAGEN) and quantified with Picogreen. Library preparation for Illumina sequencing (HiSeq 2000) was carried out using a described protocol (Garber et al., 2012) with slight modifications (Ostuni et al., 2013). The purified DNA libraries were quantified both with a 2100 Bioanalyzer (Agilent Technologies) and Qubit (LifeTechnologies) and diluted to a working concentration of 10 nM.
GEO Accession
GSM2419205