GSM8450375: WT_ChIP-Epi_input-5.0-1; Mus musculus; ChIP-Seq
Approximately 200-500 cells per sample were harvested, washed, and lysed before MNase digestion. After adding MNase stop solution, the supernatant containing fragmented chromatin is collected. Meanwhile, the target protein antibody is first incubated with Dynabeads Protein A/G (Life Technologies) for 4 hours at 4℃. Then, the supernatant is added to the antibody-beads mixture and incubated overnight 4℃. The complex is washed, and bound DNA is eluted using ChIP elution buffer. DNA purification is performed using phenol chloroform extraction and ethanol precipitation. Sequencing libraries were prepared using the KAPA Hyper Prep Kit (KAPA, KK8504, Switzerland) according to the manufacturer's instructions.