An improved simple method for DNA extraction from fungal mycelia

Abstract

A simple and inexpensive method for extraction of genomic DNA from mycelia of filamentous fungi was developed and the standardization of the protocol is described. The protocol includes both mycelium culturing and DNA extraction. By incorporating a selected commercial sand product into the culture media, the collection and the grinding of mycelia was simple and quick. The protocol of DNA extraction was derived from the alkaline lysis method for plasmid DNA extraction and modified to facilitate efficient extraction of fungal genomic DNA. It has increased efficiency and fidelity compared to the other simple DNA extraction protocols. Relative to a commonly used commercial DNA extraction kit, this extraction protocol is faster, easier and can generate more DNA without compromising quality. From a single sample of mycelium, genomic DNA can be prepared within 20 min, including concentration measurement. The DNA can be used in restriction enzyme digestion and as a template in PCR to amplify DNA fragments larger than 3000 base pairs. This method is recommended for DNA extraction in fungal phylogenetic studies when hundreds or thousands of isolates from one or a few closely-related species are subjected to PCR-based genotyping.