GSM3759784: Zfp281-ChIP_WT-ESC_I; Mus musculus; ChIP-Seq
Cells were fixed with 1% formaldehyde and neutralized with glycine to a final concentration of 0.125 mM. After nuclei extraction, DNA for ChIPs of endogenous proteins and corresponding inputs was sonicated using Diagenode Bioruptor Plus in 15 ml Diagenode Falcon tubes, cell lysates incubated with corresponding antibody over night at 4°C and Dynabeads were added the next day. In case of Bio-Ehmt1 and corresponding inputs, DNA was digested with Micrococcal Nuclease (Cell Signaling) and incubated with M-280 Streptavidin coupled Dynabeads (Invitrogen) for 1h at 4°C. Beads were washed, eluted and the suspension was de-crosslinked over night at 65 °C. DNA of ChIPs from endogenous proteins and corresponding inputs was purified using MinElute PCR Purification Kit (QIAGEN) and of Bio-Ehmt1 ChIPs and corresponding inputs using AMPure XP beads (Beckman Coulter). ChIP-seq libraries were prepared using NEBNext Ultra kit (New England BioLabs) following the manufacturer's recommendations.
GEO Accession
GSM3759784