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A simple and efficient method for extracting S. Rolfsii DNA for PCR based diversity studies

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Title A simple and efficient method for extracting S. Rolfsii DNA for PCR based diversity studies
 
Creator Ssekamate, Allan Male
Kato, Fred
Mukankusi, Clare
 
Subject exopolysaccharides
dna sequences
pathogens
exopolisacáridos
secuencia de adn
organismos patógenos
 
Description 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 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 amplification using inter-simple sequence repeat primers and primers targeting the internal transcribed spacer region of S. rolfsii.
Our method will be very useful in laboratories that don’t have access to liquid nitrogen and freeze-drying facilities and will be a catalyst for PCR-based phylogenetic studies of this important pathogen of common bean.
 
Date 2018-01-17
2022-12-06T14:05:51Z
2022-12-06T14:05:51Z
 
Type Journal Article
 
Identifier Ssekamate, A.M.; Kato, F.; Mukankusi, C.M. (2018) A simple and efficient method for extracting S. Rolfsii DNA for PCR based diversity studies. Journal of Plant Pathology & Microbiology 3 p. ISSN: 2157-7471
2157-7471
https://hdl.handle.net/10568/125802
https://doi.org/10.4172/2157-7471.1000441
 
Language en
 
Rights CC-BY-4.0
Open Access
 
Format 3 p.
application/pdf
 
Publisher OMICS Publishing Group
 
Source Journal of Plant Pathology & Microbiology