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

  • C Niu
  • H Kebede
  • DL Auld
  • JE Woodward
  • G Burow
  • RJ Wright
Keywords: CTAB, SDS, DNA isolation, phenol/chloroform free

Abstract

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 nonphenol/ 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.
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eISSN: 1684-5315