title: GSM1923444: ChIP-seq_H3K27Ac_NKT17_3; Mus musculus; ChIP-Seq
library_construction_protocol: ChIP-seq was performed as described in  Seumois et al., Nat Immunol. 2014, PMID:24997565. Briefly: frozen cells were lysed in 120 μl of lysis buffer (50 mM Tris-HCl pH 8.0, 10 mM EDTA, 1 % SDS, 1 mM phenyl methane sulfonylfluoride, 20 mg/ml sodium butyrate, proteinase inhibitor cocktail; Sigma), and chromatin was sheared by sonication, using BioRuptorTM (Diagenode), to generate 100–500 bp DNA fragments. Chromatin was diluted in 1 ml of radioimmunoprecipitation assay (RIPA) buffer and immunoprecipitation was done overnight at 4 °C by incubating 1 μl of H3K27ac antibody (rabbit polyclonal IgG, lot# Gr167929-1, Abcam) pre-coated on 10 μl protein A coated magnetic beads (Invitrogen). Immunocomplexes were captured, washed, and ChIP DNA eluted as described previously. One ng of ChIP DNA was PCR amplified for 18 to 22 cycles using whole genome amplification (WGA) primers (WGA-SEQX, Sigma) following the manufacturer’s instructions. Approximately 100 ng amplified DNA was used for preparing a standard Illumina sequencing library (TruSeq® Nano DNA HT Sample Preparation Kit, Illumina). Whole Genome Amplification (MDA); Libraries were sequenced on the HiSeq2500 Illumina platform to obtain 50-bp single end reads (TruSeq® Rapid Kit, Illumina).
attributes.tag: GEO Accession
attributes.value: GSM1923444