Journal of Plant Pathology & Microbiology

Male AS

Publications

• Research Article
  A Simple and Efficient Method for Extracting S. Rolfsii DNA For PCR Based Diversity Studies
  Author(s): Male AS, Kato F and Mukankusi CMMale AS, Kato F and Mukankusi CM
  
  Present methods of extracting DNA from Sclerotium rolfsii use a lot of hazardous organic chemicals to extract high quality DNA. Extraction of the DNA is further complicated by exopolysaccharides that bind to the DNA making it mucilaginous. We developed a simple and efficient protocol for extracting DNA high quality from S. rolfsii. Our method uses a DNA extraction buffer that contains sodium dodecyl sulphate and proteinase K to inactivate proteins and high salt concentration to precipitate the exopolysaccharides. It uses neither phenol, chloroform nor isoamyl alcohol during the DNA extraction process. It also does not require freeze drying of the mycelia and grinding in using liquid nitrogen. Using our method, a sufficient amount of pure (mean A260: A280=1.91 ± 0.001) DNA (mean = 55.57 ± 0.002 ng/µl) was obtained from 100 mg of mycelia. The DNA was amenable to PCR .. View More»
  DOI: 10.4172/2157-7471.1000441

  Abstract PDF