Peqgold fungal dna mini kit

Fungal mycelia were vacuum-filtered through sterile 70-mm-diameter filter discs MN 615 (Macherey-Nagel, Düren, Germany) and washed twice with sterile tap water. Extraction of DNA from 100 mg mycelia was performed using the peqGOLD Fungal DNA Mini kit (VWR International, Erlangen, Germany) according to the manufacturer’s instructions. Extraction of DNA from agarose gels was performed using the MinElute Gel Extraction kit (QIAGEN, Hilden, Germany) according to the manufacturer’s instructions. Genomic DNA was eluted and suspended in filtered, sterile, deionized water for all extraction protocols (Filtropur S 0.2 μm, Sarstedt, Nümbrecht, Germany). DNA concentrations were determined with a NanoDrop 1000 spectrophotometer (PeQlab, Erlangen, Germany).

PCR grade genomic DNA was prepared from 30 mL liquid cultures of the 217 isolates grown under conditions that were optimum for the respective species. Mycelia were vacuum filtered and washed twice with sterile tap water. Fifty milligram of mycelia were treated for 1 min in a Fastprep® 24 system with addition of sterile glass beads (0.4 g of 0.5 mm diameter and 0.22 g of 1.25–1.65 mm diameter) at an amplitude of 6 m/s in lysis buffer PL2 provided with the peqGold Fungal DNA Mini Kit (VWR International, Erlangen, Germany). DNA purification from treated mycelia was performed according to the manufacturer´s protocol. DNA was maintained frozen at − 20 °C until used and further stored at 4 °C after initial thawing.

Plasmid DNA was extracted using the PeqGOLD plasmid miniprep kit (VWR International GmbH, Darmstadt, Germany) according to the manufacturer’s manual.
F. culmorum was grown in PDB at room temperature with constant shaking for 3 days. Mycelia were collected, and DNA was isolated using the PeqGOLD fungal DNA mini kit (VWR International GmbH, Darmstadt, Germany) according to the manufacturer’s manual. Alternatively, DNA was extracted from fungal mycelium collected from PDA plates, in accordance with Liu et al. [41 (link)].
Fungal mycelia, as well as wheat leaves, were homogenized using a FastPrep-24™ classic instrument (MP Biomedicals GmbH, Eschwege, Germany) with liquid nitrogen and sterile steel spheres for RNA extraction. RNA isolation was performed using a plant RNA isolation mini kit (Agilent Technologies, Inc., Wilmington, NC, USA.) according to the manufacturer’s manual.