GSM8207973: 7-WT-cr; Mus musculus; OTHER
Clarify the lysate from the nucleus treated with Protein A-Tn5 enzyme and isolate the histone DNA complex with antibodies. Hyperactive Universal CUT&Tag Assay Kit for Illumina Pro (Vazyme, China) was used to perform the CUT&Tag assay. Briefly, a total of 50 000 cells were harvested and enriched by ConA magnetic beads. The samples were incubated with primary H3K18ac or H3K18cr antibody at 4°C overnight and secondary antibody at room temperature for 1 hour. Cells were incubated with the protein A-Tn5 transposon at room temperature for 1 hour, and washed thrice in Dig-300 buffer. The cells were resuspended in 50 μL of fragmentation buffer, incubated at 37 °C for 1 hour and then terminated with 1 μL of 10% SDS at 55 °C for 10 min. DNA Extract Beads Pro was used to extract the ~250bp DNA fragments. The libraries were used for sequencing on an Illumina NovaSeq 6000 platform at Novogene Science.