title: GSM9210280: BrM_IgG; Homo sapiens; OTHER
library_construction_protocol: 1 × 105 MDA-MB-231BrM cells were collected,for the CUT&Tag analysis, we utilized the Hyperactive Universal CUT&Tag Assay Kit for Illumina (TD903, Vazyme) For the CUT&Tag analysis, we utilized the Hyperactive Universal CUT&Tag Assay Kit for Illumina (TD903, Vazyme). Briefly, 1 × 105 MDA-MB-231BrM cells were collected, resuspended in 100 μL wash buffer, transferred to an 8-tube strip containing activated ConA beads, and incubated for 10 min. Subsequently, 50 μL prechilled antibody buffer containing ADAMTS1 antibodies was added to cells and incubated at 4 °C overnight, followed by the addition of diluted secondary antibodies and incubation for 60 min. After washing thrice with DIG wash buffer, samples were incubated with diluted pA/G-Tnp transposase for 1 h. Next, 50 μL diluted TTBL was added, and samples were incubated at 37 °C in a PCR instrument for 60 min for fragmentation. Subsequently, DNA extraction was performed by adding 5 μL proteinase K, 100 μL buffer L/B, and 20 μL DNA extract beads on the fragmented samples and incubating at 55 °C for 10 min. Following washes with WA and WB buffers, samples were eluted using sterile ultrapure water. The extracted DNA underwent library amplification using the TruePrep Index Kit V2 for Illumina (TD202, Vazyme). PCR products were treated with VAHTS DNA clean beads (N411, Vazyme) for 5 min, the supernatant then was discarded, samples were washed with 80% ethanol, eluted using sterile ultrapure water, and the library was sequenced on the Illumina NovaSeq platform.