2.2. Genomic DNA Extraction

Up to 200 mg of an obtained colony were harvested for gDNA extraction by adapting the salt extraction protocol described by Lohnoo et al. [21]. A hyphal mat was transferred to a 2-mL sterile plastic screw-cap tube containing 1000 mg of glass beads (710–1180 mm in diameter; Sigma, St. Louis, MO, USA) and combined with the salt homogenizing buffer (0.4 M NaCl, 10 mM Tris-HCl pH 8.0, and 2 mM EDTA; 400 μL buffer per 100 mg hyphae). To remove carry-over culture agar from the harvested hyphae, the sample tube was boiled at 100 °C for 5 min. The hyphal mat was ruptured by a Tissue Lyzer Retsch MM301 (setting: 2 min at 30 Hz; Qiagen, Hilden, Germany) and mixed with 45 μL of 20% SDS and 8 μL of 20 mg/mL proteinase K, before an overnight incubation at 56 °C. After well-mixed with 0.3 mL of 6 M NaCl, the sample was centrifuged at 10,000× g for 30 min. The supernatant was collected, combined with an equal volume of isopropanol, stored at −20 °C for 1 h, and centrifuged (10,000× g) at 4 °C for 20 min. After discarding the supernatant, a resulting pellet was collected, washed with 70% ethanol, air dried, and resuspended in 100 μL of Tris-EDTA (10 mM Tris, 1 mM EDTA; pH 8.0). A NanoDrop 2000 spectrophotometer estimated DNA concentration at 260/280 nm wavelengths (Thermo Scientific, Waltham, MA, USA).