GSM7785375: NPM1-RARG; Homo sapiens; OTHER
Cells were harvested, counted and  were washed with  Wash Buffer . Concanavalin A Beads were prepared with Binding buffer and 10 µL of activated beads were added per sample, invert to mix and incubate at room temperature . The primary antibody incubation was performed on a rotating platform overnight at 4 °C. Beads were washed and incubated with secondary antibody . After incubation, the beads were washed and incubated with hyperactive pA/G-transposon (pA/G-Tnp) at RT. Next, cells were re-suspended in  Trueprep Tagment Buffer L and  the DNA fragments were extracted with  DNA Extract Beads .  Beads were washed twice with Buffer WA and Buffer WB. After drying ~ 5 min, 20 µL ddH2O was added. To amplify libraries, 15 µL DNA was mixed with 25 µL 2×CUT&Tag amplification mix, 5 µL N5, and 5 µL N7 primer, using a different barcode for each sample. The sample was placed in a Thermocycler with a heated lid using the following cycling conditions: 72 °C for 3 min; 95 °C for 30 min; 14 cycles of 98 °C for 10 s and 60 °C for 5 s; final extension at 72 °C for 1 min and hold at 4 °C. Post-PCR clean-up was performed by adding 100 µL VAHTS DNA Clean Beads (Vazyme, #N411), and libraries were incubated with beads for 5 min at RT, washed twice gently in 80% ethanol, and eluted in 20 µL ddH2O.