title: GSM8857489: CutRun_MCF7_KO_H3K4me3; Homo sapiens; OTHER
library_construction_protocol: 200,000 cells were harvested per reaction. Permeabilized cells, bound to Concanavalin A-coated beads, were mixed with H3K4me3 antibody (CST #9751) at a dilution of 1:50, and incubated overnight at 4°C while rotating. pAG-MNase enzyme (generated in the Department of Life Sciences Core Facilities, WIS, using Addgene plasmid 123461) was added to each sample for 1 hour at 4°C. Targeted digestion was conducted for 15 minutes at 0°C under low salt conditions. DNA purification was done using Nucleospin gel and PCR clean-up kit (Machery-Nagel, 740609). Libraries were quantified by Qubit (ThermoFisher Scientific) and proper fragment range (200-400bp) was verified by TapeStation (Agilent). Sequencing was done on a Next-Seq 550 instrument (Illumina) using a Nextseq 150 cycles mid output kit, allocating 10M reads per sample (paired end sequencing).