title: GSM8577399: BV173_parental_DMSO_IgG_ChIP; Homo sapiens; ChIP-Seq
library_construction_protocol: ChIP-Seq for H3K27ac was done as described in Boulianne et al using anti-Histone H3 (acetyl K27) antibody – ChIP Grade (ab4729) (Abcam) and polyclonal rabbit IgG (isotype control) (sc2027) (Santa Cruz Biotechnologies), with the exception that initial cross-linking was performed at RT. For HA-ChIP-Seq, three lots of 2 x 10^7 cells per sample were separately, sequentially double-crosslinked consisting of a 30 min RT incubation with 2 mM Disuccinimidyl Glutarate (DSG) in PBS followed by a 30 min RT incubation with 1% formaldehyde (prepared in PBS) (rotating during incubation and washes in between), with the remainder done as described in Boulianne et al3 but using 0.8 ug anti-HA-tag antibody (Cell Signalling #3724) per 2 x 10^7 cells. ChIP-Seq libraries were generated from ChIP immunoprecipitated DNA using the NEBNext® Ultra™ II DNA Library Prep Kit for Illumina and NEBNext Multiplex Oligos for Illumina (NEB) and sequenced either at Novogene or at the in-house sequencing facility of the MRC London Institute of Medical Sciences at Imperial College London.