title: GSM5856140: E16.5_CM_p300_bioChIP-seq_rep2; Mus musculus; ChIP-Seq
library_construction_protocol: E12.5 to P42 heart ventricles were harvested, homogenized, and crosslinked in 1% formaldehyde in PBS for 10 minutes at room temperature and subsequently quenched with 125 mM glycine for 5 min. Chromatin was fragmented using a microtip sonicator (Qsonica, S-3000) at 30% amplitude for 8 minutes. Biotinylated P300 and bound chromatin was pulled down by incubation with streptavidin beads (ThermoFisher Scientific, 11206D). Bead-bound chromatin was resuspended in SDS elution buffer (50 mM Tris–HCl pH 8,10 mM EDTA, 1% SDS) and incubated overnight at 65 °C to reverse crosslinking. After RNAse A and Proteinase K treatment, ChIP DNA was purified using Qiagen MinElute columns. Libraries were constructed using the KAPA HyperPrep ChIP-seq library preparation kit (Roche, cat# 07962347001).