Ser5Pol2-CR-RAD21-Neg6h-Dox12h-Rep1
CUT&RUN was performed using the CUTANA ChIC/CUT&RUN Kit (Epicypher) on RAD21-degron treated with 0.5 M NEG or dTAGV-1 for 6 hours followed by 200 ng/ml dox for 12 hours. Nuclei were extracted, bound to concanavalin A-coated beads, and incubated with specific antibodies overnight. MNase digestion was performed, and DNA was purified using SPRIselect beads. Libraries were prepared with the NEBNext Ultra II DNA Library Prep Kit and sequenced on an Illumina NovaSeq X (PE 275 bp) at an average depth of 20 million reads per sample.