GSM8634487: a_IgGs_ChIP_2_16823; Homo sapiens; ChIP-Seq
Cells were fixed with 1% formaldehyde solution during 10 min at RT, and quenched with 0.125 M glycine during 5min at RT. Nuclei were collected at 800 x g, 4 ºC for 5 min and lysed with nuclear lysis buffer (50 mM Tris-HCl pH 8.1, 10 mM EDTA pH 8, 1% SDS; supplemented with protease inhibitors cocktail). DNA was sonicated obtaining 200-500 bp fragments. Sonicated samples were 10 times diluted with dilution buffer (16.7 mM Tris-HCl pH 8.1, 167 mM NaCl, 1.2 mM EDTA pH 8, 1% Triton X-100, 0.01% SDS; supplemented with protease inhibitors cocktail). High sensitivity quality control (Qubit), DNA libraries (Bioanalyzer, Agilent Technologies) and DNA sequencing (Sanger) were performed by Genomics Unit (Centre of Genomic Regulation (CRG), PRBB, Barcelona).