title: GSM5550302: W12 cells - Brd4_rep4; Homo sapiens; ChIP-Seq
library_construction_protocol: Cells were crossed-linked with 1% formaldehyde and chromatin was isolated and sheared to 100–500 bp DNA fragments using a Bioruptor sonicator (Diagonode) on high power settings. Chromatin samples were incubated overnight at 4°C with antibody and the resulting chromatin immunocomplexes were precipitated with blocked Dynabeads Protein G (Invitrogen), subjected to multiple wash steps and the chromatin eluted in elution buffer (50 mM Tris-HCl [pH 8.0], 10 mM EDTA [pH 8.0], 1% SDS). Chromatin was reverse cross-linked overnight at 65°C in 0.2 M NaCl, followed by RNase A and proteinase K treatment, and the DNA purified using the ChIP DNA Clean & Concentrator kit (Zymo Research). Libraries were prepared using Kapa Hyper Preps (Roche) according to the manufacturer's instructions and sequenced on the Illumina HiSeq 4000 following the manufacturer's protocol.