GSM7648606: Mnase_F_fujiA; Fusarium fujikuroi; MNase-Seq
Crosslinked and quick-frozen mycelia (3 days, liquid synthetic ICI, 6 mM glutamine) was treated with MNaseI (60 U/µL, Thermo Fisher Scientific) for 4 minutes. The reaction was stopped by the  addition of 0.5 M EDTA (final conc. 40 mM EDTA). Reverse cross-linking of digested DNA was performed for at least 30 minutes at 65 °C. Subsequently,  samples were diluted and treated with proteinase K (1h and 40°C, Thermo Fisher Scientific) for protein degradation. DNA fragments were extracted  using phenol and separated on an agarose gel to check for successful MNase digestion. Fragmented DNA was cut from the gel at 150 bp and extracted using the  Monarch® DNA Gel Extraction Kit (NEB®). The quality control, library preparation and sequencing of the samples were performed by the Vienna BioCenter Core Facilities (Vienna, Austria). Library prep and sequencing was performed using poly-A enrichment kit (NEB) and Nextera Library prep kit.