title: GSM9528689: SNB19 TES Cut&Tag (anti-V5), biological replicate 3; Homo sapiens; OTHER
library_construction_protocol: CUT&Tag was performed according to the CUTANA CUT&Tag Protocol (EpiCypher). Briefly, cells were harvested to obtain single-cell suspension, nuclei were extracted, and 100,000 nuclei were used for each replicate. Nuclei were bound to activated Concanavalin A-Conjugated Paramagnetic Beads (EpiCypher) and incubated overnight at 4C with primary antibodies: anti-V5 (mouse, Thermo Fisher Scientific R96025) for TES samples, anti-HA (rabbit, Cell Signaling Technology 3724S) for TEAD1 samples, mouse IgG (Diagenode C15400001) and rabbit IgG (Diagenode C15410206) as controls. Following overnight primary antibody incubation, nuclei suspensions were incubated with secondary antibodies (Goat Anti-Mouse IgG, Vector Laboratories I-9200-1.5; Goat Anti-Rabbit IgG, Vector Laboratories AI-1000-1.5), washed, and chromatin tagmentation was performed using pA-Tn5 Transposase (Diagenode C01070001). DNA was released from nuclei, and sequencing libraries were amplified using single-index barcodes, cleaned up, and paired-end sequenced on an Illumina NovaSeq platform.