GSM9108938: CTCF-GFP-AID mESC, NanoTag, CTCF, rep2; Mus musculus; OTHER
400,000 fresh mESCs were collected by treatment with StemPro Accutase at 37dC for 5 min and centrifugation at 600xg for 3 min and washed with Wash buffer, bound to conA beads and then incubated with either primary Ab, then secondary Ab, then pAG-Tn5 for CUT&Tag or nanobody-Tn5 for NanoTag. Tagmentation was performed at 37dC for 1 hr. Libraries were prepared using the NEBNext PCR master mix and i5 and i7 indexes for CUT&Tag. Libraries were amplified with the amount of cycles determined by qPCR required to reach 1/3 of the maximuml fluorecence. Double-sided cleanup of libraries wwas performed using AMPUre XP beads. Libraries were quantified using Agilent High Sensitivity DNA Tapestation kit.