title: GSM8190238: Chd7_KO H3K27ac rep1; Mus musculus; OTHER
library_construction_protocol: CUT&Tag library preparation was performed using percoll-purified tumor cells according to the online protocol from EpiCypher with minor modifications. Briefly, nuclei from 1 x 105 cells were used in each reaction. Of note, wheat germ agglutinin beads (BP530, Bangs Laboratories) were used instead of Concanavalin A beads to absorb the nuclei. After the binding of primary antibodies and secondary antibody (EpiCypher), targeted chromatin tagmentation was achieved by CUTANA pAG-Tn5 (EpiCypher). Reaction was terminated by adding 1.66 μl of 0.5M EDTA, 0.5 μl of 10% SDS and 0.42 μl of 20 mg/ml Proteinase K directly into each reaction, followed by incubation at 55℃ for 1 hour. Then, DNA fragments were purified by Zymo DNA Clean & Concentrator-5 kit (D4014, Zymo). Transposed DNA fragments were amplified by a 2-step PCR amplification as described in ATAC-seq library preparation. Of note, the final concentration of primers used in library PCR was adjusted to 400 nM