Full Length Research Paper

A safe inexpensive method to isolate high quality plant and fungal DNA in an open laboratory environment

Chen Niu¹, Hirut Kebede^{1, 2}, Dick L. Auld¹, Jason E. Woodward^{1, 3}, Gloria Burow⁴, and Robert J. Wright^{1, 2}*

¹Texas Tech University, Lubbock, TX, USA.

²Texas AgriLIFE Research, Lubbock, TX, USA.

³Texas AgriLIFE Extension, Lubbock, TX, USA.

⁴USDA ARS, Lubbock, TX, USA.

*Corresponding author. E-mail: robert.wright@ttu.edu. Fax: +1 806 742 0775. Tel: +1 806 742 4764.

Abbreviations: AFLP, amplified fragment length polymorphism; CTAB, cetyl-trimethyl ammonium bromide; SDS, sodium dodecyl sulfate.

Accepted 10 July, 2008

The most commonly used plant DNA isolation methods use toxic and hazardous chemicals (phenol, chloroform), which require special equipment to minimize exposure and may limit their use in certain environments. Commercial DNA extraction kits are convenient and usually safe, but their availability to certain developing countries and high cost can be limiting, especially when handing a large number of samples and considering experiments with limited financial resources. Current reports on non-phenol/chloroform protocols have not thoroughly examined the quality and suitability of the DNA for studies that require high precision. A simple, economical and rapid method is presented to isolate high quality DNA from plant and fungal species. This method uses potassium acetate to remove proteins and polysaccharides in an SDS extraction buffer. Further DNA purification is achieved using a low salt CTAB treatment. This SDS/CTAB protocol was used to isolate high quality genomic DNA subject to restriction endonuclease digestion and AFLP analysis from both plant and fungi with minimum cost and health concerns.

Key words: CTAB, SDS, DNA isolation, phenol/chloroform free.