GSM7427793: Susceptible_32 [AATB32]; Mus musculus; OTHER
Isolated nuclei was incubated with respective antibodies followed by DNA fragmentation with Micrococcal nuclease incubation. DNA was isolated using the NucleoSpin Gel and PCR Clean-Up kit CUT&RUN libraries were prepared using the NEBNext Ultra II DNA Library Prep Kit complemented with NEBNext® Multiplex Oligos for Illumina. 15nM of DNA was used as starting material for IgG and IP samples. Libraries for H3K27me1 were amplified using 13 cycles on the thermocycler. Libraries for H3K27me2 were amplified using 14 cycles on the thermocycler. Post amplification libraries were size selected at 250-350bp in length using AMPure XP beads from Beckman Coulter. Libraries were validated using the Agilent High Sensitivity DNA Kit.