GSM3414981: RH4 DMSO AQuA-HiChIP; Homo sapiens; Mus musculus; OTHER
Nuclei were then pelleted, sonicated (11 minutes shearing 'on' time with 30 seconds 'on' 30 seconds off, using Active Motif Epi-shear probe sonicator at 30% power), and immunoprecipitated with cross-species reactive antibody against H3K27ac (Active Motif, Cat# 39133). After overnight incubation (rotating, 4 °C), the ChIP reactions were buffer exchanged into Dynabeads Protein A (ThermoFischer Scientific, Cat# 10002D) and incubated (2 hours, 4 °C).  ChIP reaction was then washed, eluted, treated with Proteinase K (45 minutes at 55 °C), and crosslinks reversed by increasing temperature to 67 °C for 2 hours, and purified. Biotin capture and washing was performed with Dynabeads M-280 Streptavidin (ThermoFischer Scientific, Cat# 11205D), followed by on-bead library preparation (end repair, A-tailing, adapter ligation and library amplification)31.  Amplified library DNA was purified with AMPure beads, quantified and multiplexed, and paired-end sequenced with Illumina NextSeq High Output v2 kit (150 cycles, Cat# FC-404-2002) to generate 100-200 million read pairs per condition.
GEO
GSM3414981