anti-H3K4me1 antibody (CMA302 provided by Dr. H. Kimura)
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Trypsinised feeder-free ES and TS cells (1×10^6) were collected and fixed with 1% formaldehyde at room temperature. Crosslinking reaction was quenched with 125 mM glycine for 5 min. The cells were resuspended in SDS lysis buffer (ChIP Reagent, Nippon Gene Co., Ltd.), and the lysate was sonicated to fragment chromatin using a Covaris S220 (Covaris, Woburn, MA, USA). The sonicated lysate was subjected to immunoprecipitation with a monoclonal antibody against TEDA4, KLF5, H3K4me1, or H3K27ac. The immunoprecipitated and reverse-crosslinked DNA was purified using AMPure XP beads (Beckman Coulter, Inc., Pasadena, CA, USA) according to the manufacturer's instructions. ChIP or input DNA (0.1 ~ 1 ng) was subjected to end repair, dA-tailing, and adaptor-ligation, and amplified by 6-13 cycles of PCR.