GSM8112459: H4K16ac-con 1; Mus musculus; OTHER
Oocytes were incubated in 50 µL antibody buffer; 0.5 μg antibody was added and the mixture was incubated at 4℃ overnight. After washing twice with dig-wash buffer, 50 µL dig-wash buffer with 0.2 µg secondary antibody was added and the mixture was incubated at room temperature for 1 h. After washing twice with 100 µL dig-wash buffer, 1 µL pG/A-Tnp and 49 µL dig-300 buffer were added, and the samples were incubated at room temperature for 1 h. Next, the samples were washed twice with 100 µL dig-wash buffer. We subsequently added 200 µL tagmentation buffer and incubated the samples at 37°C for 1 h.The reaction was stopped with 5 μl Proteinase K、100 μl Buffer L/B, and extraction with DNA Extract beads. CUT&Tag was performed using the Hyperactive Universal CUT & Tag Assay Kit for Illumina (TD903, Vazyme).Oocytes were incubated in 50 µL antibody buffer;    0.5 μg antibody was added and the mixture was incubated at 4℃ overnight.After washing twice with dig-wash buffer, 50 µL dig-wash buffer with 0.2 µg secondary antibody was added and the mixture was incubated at room temperature for 1 h. After washing twice with 100 µL dig-wash buffer, 2 µL pA/G-Tnp and 49 µL dig-300 buffer were added, and the samples were incubated at room temperature for 1 h. Next, the samples were washed twice with 100 µL dig-wash buffer.We subsequently added 200 µL tagmentation buffer and incubated the samples at 37°C for 1 h. The reaction was stopped with 5 μl Proteinase K、100 μl Buffer L/B; After extraction with DNA Extract beads PCR was performed to amplify the libraries under the following conditions: 72°C for 3 min, 98°C for 30 s, 17 cycles of 98°C for 10 s and 60°C for 5 s, and a final extension at 72°C for 1 min with a hold at 4°C.