GSM3936986: ChIP_MB_H3k27ac_repeat 2; Mus musculus; ChIP-Seq
Lysates were clarified from sonicated nuclei and proteine-DNA complexes were isolated with antibody. Dissociated tissues were fixed for 15 min at room temperature with 1% formaldehyde-containing medium. Nuclei were isolated and sonicated in sonication buffer (10 mM Tris-HCl pH 8.0, 1 mM EDTA, 0.5 mM EGTA and protease inhibitor cocktail). Sonicated chromatin (~300 µg) was used for immunoprecipitation by incubation with appropriate antibodies (4 mg) overnight at 4°C. Ten percent of chromatin used for each ChIP reaction was kept as input DNA. Pre-rinsed magnetic protein A/G beads (50 ml) were added to each ChIP reaction and reactions were incubated for 1 hr at 4°C.The beads were then incubated in 200 ml elution buffer at 65°C for 20 min to elute immunoprecipitated materials. The ChIP-seq libraries were prepared using NEBNext ChIP-seq Library Prep Master Mix Set for Illumina (NEB catalogue number E6240L) and then run on the Illumina sequencer HiSeq 2500.
GEO Accession
GSM3936986