Neurospora crassa MNase (Micrococcal nuclease) seq

Micrococcal nuclease (MNase) experiments was performed as previously described(Seymour et al., 2016). Neurospora was cultured in petri dishes in liquid medium for two days.Afterwards, the Neurospora mats were cut into discs and transferred into medium-containingflasks and were harvested at DD14. After crosslinking with 1% formaldehyde for 30 min atroom temperature, nuclei were incubated with micrococcal nuclease (Takara) for 60 min at 37 °Cbefore the reaction was stopped by addition of EDTA. Sequencing libraries were generated using50 ng of DNA purified from the MNase-digested chromatin (Diagenode DNA sample prep kit)and size-selected by agarose gel purification to ensure insert sizes close to that of DNA bound bymononucleosomes. After 75bp paired-end sequencing, the MNase-seq data were normalized bytotal number of reads.