GSM9017746: C17, CUT&RUN-seq, IgG; Homo sapiens; OTHER
The CUTANATM ChIC/CUT&RUN Kit (Cat# 14-1048) was used in this experiment. A total of 500,000 cells were harvested and bound to concanavalin A-coated (ConA) magnetic beads. The cell membranes were permeabilised with 5% digitonin, allowing the primary antibody to bind histones with posttranslational modifications (PTMs) or transcription factors (TFs). CUTANA™ pAG-MNase (Cat# 15-1016) at a volume of 2.5 µL was incubated with the samples for 10 minutes at room temperature to target the primary antibody heavy chain at the region of interest. A 100 mM calcium chloride solution was used to initiate DNA digestion for 2 hours at 4°C. The enriched DNA was purified and prepared for sequencing libraries. One to ten nanograms of enriched DNA was collected to generate the sequencing libraries using the NEBNext® Ultra™ DNA Library Prep Kit for Illumina® (Cat# E7370L). NEBNext® Ultra™ DNA Library Prep Kit for Illumina® (Cat# E7370L)