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

Chen Niu, Hirut Kebede, Dick L. Auld, Jason E. Woodward, Gloria Burow, Robert J. Wright

Research output: Contribution to journalArticlepeer-review

31 Scopus citations

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 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.

Original languageEnglish
Pages (from-to)2818-2822
Number of pages5
JournalAfrican Journal of Biotechnology
Volume7
Issue number16
StatePublished - Aug 18 2008

Keywords

  • CTAB
  • DNA isolation
  • Phenol/chloroform free
  • SDS

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