GSM5159894: Hudep2_D3_H3K27AC_HiCHIP; Homo sapiens; ChIP-Seq
HiChIP libraries of 10 × 1E+6 cells per sample using an H3K27ac antibody were prepared as previously described, with the following modification. After proximity ligation, the nuclear lysates were sonicated with a Branson 250 sonicator (microtip, 100% duty, 20% output, 12 × 10 s intervals). We used 8 µg of H3K27ac antibodies for each sample. Reverse cross-linked ChIP DNAs were purified with the DNA Clean & Concentrator-5 purification kit (Zymo Research) and quantified with Qubit (Life Technologies). The samples were then end-repaired, dATP-labeled, and adaptor-ligated with an NEBNext Ultra II DNA library prep kit, followed by streptavidin pull-down and PCR amplification of the library. PCR products (300–1,000 bp) were size selected by using E-gel (Invitrogen). Libraries were 100-bp paired-end sequenced with an Illumina HiSeq 4000 to obtain approximately 300 × 106 reads.