title: GSM4200146: Hmga2-/- Input; Mus musculus; ChIP-Seq
library_construction_protocol: Cells were cross-linked with 1% formaldehyde for 10 min at room temperature. The reaction was quenched for 5 min on ice by adding 0.125 M glycine. After three washes with 1x PBS, cells were subjected to hypotonic buffer L1 (50 mM Tris-HCl pH 8.0, 2 mM EDTA pH 8.0, 0.1% IGEPAL CA-630 (Sigma), 10% glycerol, 2 mM DTT, 10 mM NaF, 2 mM Na3VO4, 0.2 M imidazole, 1 mM PMSF and protease inhibitor cocktail). Cells were lysed, and sonicated with Diagenode Bioruptor to an average DNA length of 100-300 bp. After centrifugation, the soluble chromatin was immunoprecipitated using antibodies specific against H3 (Abcam), H2A.X (Millipore), pPolII (Abcam) and GADD45A (Santacruz). Precipitated chromatin complexes were removed from the beads and decrosslinked by incubating with 100 μl of 10 mM Tris-HCl pH 8.0, 1 mM EDTA pH 8.0, 0.1 M NaHCO3, 1% SDS and Proteinase K at 65 °C overnight. Reverse cross-linked immunoprecipitated chromatin was purified using the QIAquick PCR purification kit (Qiagen). Libraries were prepared according to Illumina's instructions accompanying the Ovation Ultra Low Kit.
attributes.tag: GEO Accession
attributes.value: GSM4200146