GSM9064928: MCF7-ABER cells, CtrASOM_1, H3K9me3, repeat1; Homo sapiens; OTHER
MCF7-ABER cells were collected from cultures.The cells were then incubated with ConA Beads Pro (Vazyme, China, cat. TD904) for 10 minutes at room temperature to facilitate cell attachment. Following incubation, a 1:50 dilution of the primary antibody against H3K9ME3 protein (Active Motif, cat. 39062) was added to the cells. The mixture was gently vortexed and subjected to a brief centrifugation to ensure even distribution of the antibody. The cells were then incubated with the primary antibody overnight at 4°C to allow for specific protein binding. The following day, the appropriate secondary antibody was added at a 1:100 dilution, and the cells were incubated on a rotating platform at room temperature for 60 minutes. After incubation, the cells were washed twice using a magnetic rack to remove any unbound secondary antibodies present in the supernatant.then, the cells were resuspended, and a 1:50 dilution of pA/G-Tnp Pro (Vazyme, China, cat. TD904) was added to the cell suspension. The mixture was gently mixed and centrifuged to promote binding. The binding reaction with pA/G-Tnp Pro was carried out at room temperature for 1 hour on a rotating platform. To eliminate any unbound pA/G-Tnp Pro, the cells were washed twice using a magnetic rack. The cells were then resuspended and incubated at 37°C for 1 hour to activate the transposase enzymatic activity. To halt the tagmentation reaction, 10% SDS was added, and the sample was incubated at 55°C for 10 minutes. Last, The tagmented DNA was recovered using DNA Extract Beads Pro (Vazyme, China, cat. TD904) CUT&Tag library preparation was performed using the Hyperactive Universal CUT&Tag Assay Kit for Illumina Pro (Vazyme, China, cat.TD904) according to the manufacturer instructions.