2.4.2. Preparation of Standard Curve

Material for preparation of standard curve was a series of 10-fold dilutions of DNA isolated from pure culture of researched six *Fusarium* species (*F. avenaceum, F. culmorum*, *F. graminearum, F. langsethiae, F. poae* and *F. sporotrichioides*). Pure fungal cultures were grown on PDA medium (Difco, Becton, Dickinson and Company Sparks, MD, USA) on Petri dish and DNA was isolated from scraped and lyophilized mycelium using the same protocol as for grain.

### 2.4.3. Preparation of DNA Samples for Real-Time PCR

Concentrations of DNA obtained from kernels were measured with Quantus fluorometer (Promega, Madison, WI, USA). All samples were diluted in sterile deionized water to 10 ng·μL−1. Final concentration of *Fusarium* DNA in a sample was expressed in picograms per 100 ng of total DNA.
