title: GSM6720998: Trim33KO_E2f4; Mus musculus; ChIP-Seq
library_construction_protocol: Cells were fixed with 1% formaldehyde for 5 min and quenched for 5 min with glycine. Cells were washed and resuspended in ChIP lysis buffer 1 (25 mM Tris-HCl pH 8.0, 1 % TritonX100 / 200 mM NaCl / protease & phosphatase inhibitors) and incubated 30 min on ice. Pelleted nuclei were resuspended in ChIP lysis buffer 2 (25 mM Tris-HCl pH 8.0, 1% Triton X-100, 300 mM NaCl, 0.1% SDS) and fragmented using Hilescher UP200st sonifier. Cleared lysates were incubated with E2f4 antibody overnight at 4°C on a rotating wheel. Antibodies were collected using magnetic protein A beads followed by proteinase K treatment and reversal of crosslinking. The DNA was purified by phenol/chloroform extraction and precipitated with ethanol. Sequencing libraries were prepared using NEB Ultra II DNA library Prep kit according to the manufacturer's instructions.