GSM7789944: 4CL.H3K27ac_rep3; Homo sapiens; OTHER
For CUT&Tag library construction, Hyperactive In-Situ ChIP Library Prep Kit for Illumina (Vazyme, TD902-01) was used. For CUT&Tag library construction, Hyperactive In-Situ ChIP Library Prep Kit for Illumina (Vazyme, TD902-01) was used. In brief, 100,000 cells were incubated with Concanavalin A-coated magnetic beads and then primary antibodies were added to incubate overnight at 4°C. Tubes were placed on a magnetic rack to remove the supernatants. Secondary antibodies were added and samples were incubated at room temperature for one hour. After placing the tubes on the magnetic rack and removing the supernatants, hyperactive pG-Tn5/pA-Tn5 transposon (incubated at room temperature for one hour) were added. Tagmentation was performed for 1 hour and DNA was extracted using phenol–chloroform–isoamyl alcohol. Purified DNA fragments were amplified by PCR for 15-20 cycles and the PCR products were purified following the kit's instructions.