GSM9286334: S9.6 IFN  rep1; Homo sapiens; OTHER
The CUT&Tag assay was conducted using Hyperactive Universal the CUT&Tag Assay Kit for Illumina (Vazyme, Cat No. TD903) according to the manufacturer's instructions. 80,000 cells were harvested and washed twice in wash buffer. Concanavalin A beads were then added to the cell suspension and incubated at room temperature for 10 mins. Bead-bound cells were separated from the supernatant and were resuspended in antibody buffer. Antibodies against S9.6 (Absoluteantibody, Cat No. AB01137, with 1 μg) were added to the cells, and incubated at 4°C overnight. The following day, the secondary antibody diluted in dig-wash buffer was added to the beads and incubated at room temperature for 1 h. After incubation, beads were washed three times with dig-wash buffer. pA/G-Tnp transposomes diluted in dig-300 buffer were incubated with beads at room temperature for 1 h. After three washes with dig-300 buffer, samples were then tagmented with TTBL from the kit. After 1 h of tagmentation, proteinase K and spike-in were added. Fragmented DNA was purified using DNA Extract Beads. The eluted DNA was amplified using Illumina i7/i5 indexed primers. After that, VAHTS DNA Clean Beads (Vazyme #N411) were employed for the purification of the libraries, which were then subjected to quality control and Illumina sequencing.