title: GSM6870660: adipose stromal cells, untreated - KDM6B, Rep 2 [CR_KDM6B_05]; Homo sapiens; OTHER
library_construction_protocol: After treatment, KDM6B binding was performed using the CUT&RUN methods: Briefly, Cells (1.103) were bound to Concanavalin A beads (Polysciences) and incubated with 1 μl of anti-KDM6B (Abcam, ab38113) antibodies, overnight at 4°C. pA/G-MNase (0.89 ng/μl) was bound to antibodies and activated with CaCl2 to digest chromatin during 30 min at 0°C. Enzymatic reaction was stopped and chromatin fragments were released by an incubation of 10 min at 37°C. Chromatin sonication was performed with the EpiShearTM Probe Sonicator (Active Motif). DNA was purified with MinElute PCR Purification Kit (Qiagen). Libraries were prepared by Integragen, with the CUTANA CUT&RUN Library Prep Kit and accordingly to manufacturer recommendation. Paired-end.