GSM6944434: PBMC_CTRL-1_ETS1_CutTag; Homo sapiens; OTHER
10,000 fresh cells were counted and processed according to Hyperactive® Universal CUT&Tag Assay Kit for Illumina (Vazyme, TD903) instructions. Briefly, ConA beads were added to the cell pellet and incubation at room temperature for 10 min. Subsequently, the mixed complexes were incubated overnight at 4°C with primary antibody. The following day, the cells were washed and incubated with diluted secondary antibody at room temperature for 2 h. Antibody-protein-DNA complexes were then linked by pA/G-Tnp by incubation at room temperature for 1 h. After washing, the eluted complexes were subsequently sheared by adding 5 × TTBL. The proteinase K and DNA extract beads were used for obtaining eluted DNA. Sequencing libraries were established after 11 cycles of PCR amplification. Selected size-selected were proceeded using AMPure XP beads and the DNA were sequenced on the Illumina Hiseq-PE150 platform to a depth of 2.5 × 107 reads.