CUT&Tag-seq of human cells: NPC
CUT&Tag was performed with CUT&Tag-IT Assay Kit (cat. # 53160, ActiveMotif) according to the manufacturers instructions. Cultured cells were collected and bounded to Concanavalin Acoated beads at RT for 10 min. Subsequently, cells were resuspended in antibody buffer and incubated with primary antibodies against H3K18la at 4 overnight and Guinea pig-anti rabbit IgG in Dig-wash buffer for 1h at RT in order. Samples were then mixed with pA-Tn5-adapter transposases (pA-Tn5) following by transposon activation and DNA tagmentation. After that, DNA was isolated, amplified, and purified to construct the CUT&Tag libraries for sequencing. The libraries were sequenced on an Illumina NovaSeq 6000 platform.