4mM 4-deoxypyridoxine hydrochloride in serum-free medium
chip-antibody
H3K9ac (Abcam, ab4441)
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Cells were washed once with 1X DPBS and then cross-linked. The cross-linked material was then rinsed, pelleted, and frozen in liquid nitrogen for later use. Frozen cell pellets were resuspended in lysis buffer and sonicated to shear DNA. Sheared DNA was immunopercipitated with anti-Histone H3 (acetyl K9) antibody. A negative control ChIP reaction was performed using normal rabbit IgG. The immunopercipitate was washed on protein G beads and reverse crosslinked with Proteinase K at 62 o C for 4 hours. Libraries were prepared with NEBNext® Ultra™ DNA Library Prep Kit for Illumina® (NEB) according to manufacturer instructions.