GSM8174727: HPSC,Input1; Danio rerio; ChIP-Seq
The cells were cross-linked with a final concentration of 1% formaldehyde followed by quenching with glycine. The cells were cross-linked with a final concentration of 1% formaldehyde and lysed with lysis buffer. Nuclei was sonicated to fragments about 300 - 500 bp. Dynabeads Protein A was incubated with antibody for 2-3 hours. The fragmented chromatin was transferred to the bead-antibody complex tubes and rotated at 4°C overnight. The beads were washed once with low salt buffer (10mM Tris-HCl pH7.5, 250mM NaCl, 1mM EDTA, 0.5mM EGTA, 1% Triton X-100, 0.1% SDS, 0.1% Na-deoxycholate, Cocktail proteinase inhibitor) and twice with high salt buffer (10mM Tris-HCl pH7.5, 500mM NaCl, 1mM EDTA, 0.5mM EGTA, 1% Triton X-100, 0.1% SDS, 0.1% Na-deoxycholate, Cocktail proteinase inhibitor).After crosslink reversal, library was constructed using NEBNext Ultra II DNA Library Prep Kit and sequenced using an Illumina sequencing platform.