title: GSM8010015: 4 hr DHT VCaP Input rep 2 (ARv7); Homo sapiens; ChIP-Seq
library_construction_protocol: Cells were crosslinked with fresh 1% methanol-free formaldehyde (FA) for 10 minutes. This step was carried out for immunoprecipitation (IP) of histone modification marks (H3K27Ac, H3K4me1), ARfl, FOXA1, BRD4, and p300. Double fixation: For IP of ARv7, cells were first crosslinked in 2 mM fresh disuccinimidyl glutarate for 35 minutes, followed by 1% FA for an additional 10 minutes. The samples were sonicated using the Covaris E220 in sarkosyl lysis buffer. ChIP-seq libraries for ARv7, ARfl and H3K27Ac ChIP DNA were generated using the ThruPLEX DNA-seq Kit (Rubicon Genomics) according to the manufacturer's instructions and standard 8 bp Illumina primers. Libraries were pooled and sequenced on the Illumina NextSeq 500 platform at the Molecular Biology Core Facility (DFCI). ChIP-seq libraries for H3K4me1, FOXA1 were prepared using Accel-NGS 2S Plus DNA Libraries Kit with Unique Dual Indexing (Swift Biosciences, cat# 29096/290384) according to the manufacturer's instructions. ChIP-seq libraries for H3K27Ac, BRD4 and p300 were generated using SimpleChIP ChIP-seq Multiplex Oligos for Illumina with Dual Index Primers (Cell Signaling Technology, cat# 47538) according to the manufacturer's instructions. Libraries were pooled and sequenced using a NovaSeq 6000 Sequencing System.