title: GSM7504863: PROTAC_d3NPC_K27ac; Homo sapiens; OTHER
library_construction_protocol: ESCs and NPCs were collected using TrypLE. Nuclei were isolated, fixed for 2 minutes with 0.1% Formaldehyde at room temperature, and cryopreserved prior to all Cut&Tag experiments. For each experiment, nuclei were thawed and 100,000 nuclei were used per reaction. Nuclei were bound to activated Concanavalin A beads and 0.5ug of primary antibody was bound overnight at 4C. Secondary antibody was bound for 30 minutes at room temperature, pAG-Tn5 was bound for 1 hour at room temperature, and the  tagmentation reaction was incubated at 37C for 1 hour. Libraries were amplified with NEBNext high-fidelity PCR mix, cleaned-up with AMPure XP beads, and sequenced by the NIEHS Epigenomics and DNA Sequencing Core Laboratory.