GSM9313264: CUT&RUN of H3K27me3 in MXP5 IK1 rep2; Homo sapiens; OTHER
The CUTANA CUT&RUN kit (EpiCypher #14-1048) was used according to manufacturer's instructions, using 500,000 cells per reaction. Briefly, Concanavalin A beads were used to immobilize cells and .5 μg of individual antibodies against H3K4me3 (Epicypher #13-0041), H3K27me3 (Active Motif, #39159), H3K9me3(Active Motif, #39161) or H3K36me3 (Active Motif #61101) were incubated overnight at 4°C with light rocking using  1 μL control IgG (Epicypher #13-0042) antibodies. Antibody chromatin complexes were treated with pAG-MNase for 2 hours at 4°C and DNA fragments were purified. DNA concentration was measured with a Qubit fluorometer and 5-10ng of purified DNA was input into sequencing library preparation with NEBNext UltraII kit (NEB # E7645). Library size was assessed with an Agilent Bioanalyzer and sequenced paired-end 150bp with Illumina.