title: GSM6234619: HEK293T, H3K27ac, HS2, dCas9; Homo sapiens; OTHER
library_construction_protocol: CUT&RUN assays were carried out using the Epicypher CUTANA ChIC/CUT&RUN Kit (Epicypher 14-1048). Briefly, 5.0e5 transfected cells were harvested, immobilized on concanavalin A beads, and permeabilized in 0.01% digitonin cell permeabilization buffer. The cell-bead conjugate mixture was then divided equally into 2 aliquots and incubated in 50ul antibody buffer with either 0.5µg of anti-H3K27ac antibody (Abcam, 4279) or 0.5µg of control rabbit IgG antibody (Epicypher, 14-1048) overnight at 4°C. After washing the beads, pAG-MNase was added to the immobilized cells and the solution was incubated for 2 hours at 4°C to digest and release chromatin DNA Libraries were constructed using NEBNext® Ultra™ II DNA Library Prep Kit for Illumina® kit (NEB, E7645) by Azenta Life Sciences