GSM9978522: Fep1-5FLAG, P+, 4hr; Schizosaccharomyces pombe; ChIP-Seq
The cells were cross-linked with 3% paraformaldehyde at 18℃ for 30 min, followed by secondary cross-linking with 10 mM dimethyl adipimidate at 25℃ for 45 min. Cell lysis was performed by bead beating in ChIP lysis buffer (50 mM HEPES-KOH [pH 7.5]; 140 mM NaCl; 1 mM EDTA [pH 8.0]; 1% Triton X-100; 0.1% sodium deoxycholate; and 0.1 mM PMSF) supplemented with protease inhibitors. ChIP-seq libraries were prepared using the NEBNext Ultra II DNA Library Prep Kit for Illumina and NEBNext Multiplex Oligos for Illumina with amplification using Q5 Master Mix. The DNA fragments that were 200–700 bp were enriched using AMPure XP beads.