A simple method of genomic DNA extraction suitable for analysis of bulk fungal strains.

AIMS

A simple and rapid method (designated thermolysis) for extracting genomic DNA from bulk fungal strains was described.

METHODS AND RESULTS

In the thermolysis method, a few mycelia or yeast cells were first rinsed with pure water to remove potential PCR inhibitors and then incubated in a lysis buffer at 85 degrees C to break down cell walls and membranes. This method was used to extract genomic DNA from large numbers of fungal strains (more than 92 species, 35 genera of three phyla) isolated from different sections of natural Ophiocordyceps sinensis specimens. Regions of interest from high as well as single-copy number genes were successfully amplified from the extracted DNA samples. The DNA samples obtained by this method can be stored at -20 degrees C for over 1 year.

CONCLUSIONS

The method was effective, easy and fast and allowed batch DNA extraction from multiple fungal isolates.

SIGNIFICANCE AND IMPACT OF STUDY

Use of the thermolysis method will allow researchers to obtain DNA from fungi quickly for use in molecular assays. This method requires only minute quantities of starting material and is suitable for diverse fungal species.