GSM4476409: mESC_H3K4me3_CUTTag_200; Mus musculus; OTHER
The CUT&Tag is performed according to previously described (cite CT paper) with CUT&Tag kit (Vazyme Biotech, TD091). Briefly, cells were incubated with 10μl pre-washed ConA beads (Vazyme Biotech, TD091) in 1.5ml low-binding tube. 50μl antibody buffer (Vazyme Biotech, TD091) with 0.5μg antibody was added and cultured for 2 hours at room temperature. After twice of wash with dig-wash buffer (Vazyme Biotech, TD091), 50μl dig-wash buffer with 0.5μg secondary antibody was added and incubated at room temperature for 30min. After twice of wash with 800μl dig-wash buffer, 0.58μl pG-Tn5 was added with 100μl dig-300 buffer (Vazyme Biotech, TD091). Samples were incubated at room temperature for 1 hours and then washed twice with 800μl dig-300 buffer. 300μl tagmentation buffer was added and samples were incubated at 37°C for 1 hours. The reaction was stopped with 10μl 0.5M EDTA，3 μl 10% SDS和2.5μl 20 mg/ml Proteinase K. After extraction with phenol-chloroform and ethanol precipitation, PCR was performed to amplify the libraries (Vazyme Biotech, TD094).