title: GSM5819100: PHF6-WT_CHD4_L58; Homo sapiens; ChIP-Seq
library_construction_protocol: Cells were double-crosslinked with disuccinimidyl glutarate and formaldehyde followed by lysis and nuclei isolation. Nuclei were sonicated in a Bioruptor Pico (Diagenode) and clarified chromatin was immunoprecipitated by antibodies against either histone marks or chromatin remodeling factors. Libraries were prepared according to manufacturer's instructions accompanying the NEBNext® Ultra™ II DNA Library Prep Kit for Illumina® (NEB #E7645S/L). Briefly, DNA was end-repaired followed by 5´ phosphorylation and dA-tailing. Next, adaptor ligation was carried out using NEBNext Adaptor followed by U excision using USER enzyme. Adaptor-ligated DNA was cleaned up using AMPure XP beads (Beckman Coulter, A63881), PCR amplified with NEBNext Multiplex Oligos for Illumina (Set 1, NEB #E7335; Set 2, NEB #E7500) for 8-10 cycles and PCR amplified library was cleaned up using AMPure XP beads. Purified libraries were quantified based on the average fragment size according to Bioanalyzer and Qubit results, pooled at equimolar concentrations, and sequenced as a pool to a total read-depth target of approximately 4 × 107, paired-end, 150-nucleotide-long reads per sample on an Illumina HiSeq 2500 following the manufacturer's protocols.