GSM8574672: Php3-GFP_moc3∆_FE_Rep2; Schizosaccharomyces pombe; ChIP-Seq
Cells were crosslinked using 3% PFA at 18°C for 30 min and cells were spun at 2,000g for 5 min, washed with PBS and treated with DMA at room temperature for 45min. Cells pellets were lysed using glass beads and lysis buffer (50mM Hepes/KOH, 140mM NaCl, 1 mM EDTA, 1% Triton X100, 0.1% DOC plus protease inhibitors). Cell lysates were sheared using a Bioruptor-300 (Diagenode) to an approximate size of 300–600 bp. Sequencing libraries were prepared using the NEBNext Ultra II DNA Library Prep Kit for Illumina according to the manufacturer's protocol and analyzed using an Agilent 4200 TapeStation system (Agilent).