GSM4145976: ChIP_HEC-1-B_a2_FWD_mut_rep1; Homo sapiens; ChIP-Seq
4 × 106 of cells were cross-linked by 1% formaldehyde in 10% FBS/PBS for 10 min at room temperature. Cells were then lysed twice with ice-cold lysis buffer (20 mM Tris-HCl, 2 mM EDTA, 1% Triton X-100, 0.1% SDS, 0.1% sodium deoxycholate and 1× protease inhibitors, pH 7.5) for 10 min with slow rotations. The lysed cells were then sonicated to obtain DNA fragments of about 200-500 bp using the Biorupter system (high energy, with working time of 30 seconds and resting time of 30 seconds, 30 cycles). After removal of the insoluble debris, the lysate was incubated with specific antibodies against CTCF (07-729; Millipore), RAD21 (ab992; Abcam), or H3K27ac (ab4729; Abcam) and purified by protein A-agarose beads (16-157; Millipore). NIPBL ChIP-seq were from recently published data49. ChIP DNA was extracted and prepared for high throughput sequencing using a DNA library preparation kit for Illumina (NEB). ChIP-seq libraries were sequenced on a HiSeq X Ten Platform (Illumina).
GEO Accession
GSM4145976