title: GSM8792080: Lamin A/C knockout Hela cells, target PCNA, biol rep 2; Homo sapiens; OTHER
library_name: GSM8792080
library_construction_protocol: Approximately 100,000 Hela cells were harvested and permeabilized with digitonin in wash buffer. ConA beads were activated and incubated with cells at room temperature for 10 minutes. Primary antibodies（PCNA） against the target protein were added, and the reaction was incubated overnight at 4°C. Cut&Tag was performed using the Vazyme CUT&Tag kit (#TD904) according to the manufacturer's instructions, with minor modifications. Briefly, approximately 100,000 Hela cells were harvested and permeabilized with digitonin in wash buffer. ConA beads were activated and incubated with cells at room temperature for 10 minutes. Primary antibodies（PCNA） against the target protein were added, and the reaction was incubated overnight at 4°C. The next day, samples were washed and incubated with a secondary antibody conjugated to Protein A-Tn5 transposase. Following tagmentation, DNA was extracted using DNA Clean Beads (Vazyme #N411) and purified. The libraries were amplified using the Vazyme library amplification primers (#TD202/TD203) and sequenced on an Illumina platform.