DNA extraction method for PCR in mycorrhizal fungi

Aims: To develop a simple and rapid DNA extraction protocol for PCR in myco rrhizal fungi. Methods and Results: The protocol combines the application o f rapid freezing and boiling cycles and passage of the extracts through DNA purification columns. PCR amplifiable DNA was obtained from a number of en do- and ecto-mycorrhizal fungi using minute quantities of spores and myceli um, respectively. Conclusions: DNA extracted following the method, was used to successfully a mplify regions of interest from high as well as low copy number genes. The amplicons were suitable for further downstream applications such as sequenc ing and PCR-RFLPs. Significance and Impact of the Study: The protocol described is simple, sho rt and facilitates rapid isolation of PCR amplifiable genomic DNA from a la rge number of fungal isolates in a single day. The method requires only min ute quantities of starting material and is suitable for mycorrhizal fungi a s well as a range of other fungi.