GSM5733165: ESC2; Homo sapiens; OTHER
Cells (1x10^5) were counted and centrifuged at 500 x g for 5 min at room temperature. The CUT&Tag assay was performed as described previously (Kaya-Okur, Wu et al., 2019). Briefly, cells were bound to concanavalin A-coated magnetic beads and permeabilized through the cell membrane by digitonin. The enzyme pA-Tn5 transductase binds precisely to DNA sequences near the target protein under the guidance of antibodies. The enzyme pA-Tn5 transglutaminase binds precisely to DNA sequences near the target protein under the guidance of BAF45D antibodies, thus enabling the targeted labeling of factors. DNA sequences are labeled, and connectors are added simultaneously at both ends, which can be enriched by PCR to form a sequencing-ready library. The library was enriched by PCR to form a sequencable library. After PCR, the library was purified with AMPure beads, and the library quality was assessed on an Agilent Bioanalyzer 2100 system.