Strategically eliminating damage caused by fungus requires precise identification of the fungal species present (using either conventional or molecular diagnostic techniques). The first step in performing molecular diagnostics on wood-decay fungi is to obtain analyzable fungal DNA.
Five commercially available kits and one CTAB method were tested and analyzed in order to determine effective methods for extracting DNA from field samples. The manual NucleoSpin® Plant II Kit (Macherey-Nagel, MN) yielded the best results, both in terms of the quality and quantity of the fungal DNA extracted from wood samples .
Modern diagnostic methods such as chip technology, however, require contamination-free and automated systems that can handle a large sample throughput in a short period of time. When used with appropriate kits for plants, yeasts and bacteria, currently available automated extraction systems yield fungal DNA in smaller quantities and poorer quality than manual extraction methods, thereby limiting the suitability of the former.
To overcome these difficulties, we worked with the manufacturer of the InnuPure® C16 automated extraction system (Analytik Jena, AJ) to adjust and optimize individual work sequences according to both the problem at hand and the type of field samples to be analyzed. Our aim was to achieve results that would be comparable to the preferred manual kit for extracting fungal DNA from wood samples.
Effective DNA extraction method for molecular diagnostics on wood-decay fungi