GSM7780713: PC-3 cells-PHF8; Homo sapiens; OTHER
CUT&Tag assay was performed according to its manufacturer's instructions (Hyperactive® Universal CUT&Tag Assay Kit for Illumina, Vazyme, Cat# TD903). Cells were trypsinized and centrifuged at 1000 r.p.m for 5 minutes. Then cells and beads were suspended with wash buffer and binding buffer respectively. Cells were incubated with beads for 20 minutes at room temperature. Then, 3 μg of primary antibody [PHF8 (Cell Signaling Technology Cat# 93801, RRID: AB_2800215) and E2F1 (Abcam Cat# ab179445)] was added and incubated overnight at 4 °C with slow rotation. After removing primary antibodies, secondary antibodies were added and incubated for 1 hour at room temperature. Dig-wash buffer, pA/G-Tnp, Dig-300 buffer and 5XTTBL were successively used for fragmenting DNA. Then DNA fragments were purified and extracted with Proteinase K, 100 μl Buffer L/B和20 μl DNA Extract Beads. DNA library was constructed with proper primers and amplified for 20 cycles (TruePrep® Index Kit V2 for Illumina, Vazyme, Cat# TD202).