title: GSM6166515: mycelia - H3K4me3_set1_rep2_IP; Ustilaginoidea virens; ChIP-Seq
library_construction_protocol: After the cross-linking was stopped with 125 mM glycine, the mycelia were ground in liquid nitrogen and suspended with the nuclei isolating buffer (10 mM Tris pH 8.0, 10 mM sodium butyrate, 400 mM sucrose, 0.1 mM PMSF, 5 mM β-mercaptoethanol and 1 × proteinase inhibitors). The precipitated nuclei were treated with 1 mL lysis buffer (50 mM HEPES pH 7.5, 150 mM NaCl, 1 mM EDTA, 10 mM sodium butyrate, 0.1% deoxycholate, 1% Triton X-100, 0.1% SDS, 1mM PMSF and 1 × Roche protease inhibitor cocktail) and broken into DNA fragments between 200 -500 bp using Diagenode Vioruptor. After pretreating with 10 μL protein A beads (Thermofisher, 10001D), the supernatant was incubated with anti-H3K4me3 antibody (Abcam, ab1012) for 1 h. Then 20 μL protein A beads were added into the above reaction system to bind anti-H3K4me3 antibody. Two biological repeats were conducted. The purified DNA was used as libraries construction with the NEBNext Ultra II DNA Library Prep Kit for Illumina (NEB, E7645L).