title: GSM4782224: VCAP_H3K27ac_HiChIP_DHT_24hr; Homo sapiens; OTHER
library_construction_protocol: 10 million cells were crosslinked in 1% formaldehyde for 10 minutes at room temperature and then quenched by 125mM glycine. After washing in PBS, the crosslinked cells were lysed in Hi-C lysis buffer (10 mM NaCl, 10 mM Tris-HCl, 0.2% NP-40 and 1×protease inhibitor) and digested by MboI restriction enzyme (NEB-R0147). The biotin-dATP (Thermo 19524016) was incorporated to DNA in a fill-in master mix containing DNA polymerase I (NEB-M0210). After ligation and sonication, the sheared chromatin was incubated together with Protein A beads and anti-H3K27ac antibody (Abcam, ab4729) for overnight at 4°C with rotation. ChIPed DNA was purified from the eluted chromatin using Zymo DNA Clean & Concentrator kit. Biotin-labeled DNA was captured by Streptavidin C-1 bead (Thermo Fisher, Cat#65002) and fragmented by Tn5 enzyme to generate library for high throughput sequencing. Next generation sequencing was conducted on a HiSeq4000 machine with 150 bp pair-end reads. Libraries were prepared using the Tn5 transposase. HiChIP