期刊
CANADIAN JOURNAL OF PLANT PATHOLOGY
卷 32, 期 3, 页码 396-401出版社
TAYLOR & FRANCIS INC
DOI: 10.1080/07060661.2010.508620
关键词
DNA isolation; dot-blot hybridization; mycelia; protocol; PCR
资金
- Alberta Canola Producers Commission
- Manitoba Canola Growers Association
- SaskCanola and the Canola Council of Canada
- Alberta Crop Industry Development Fund
A rapid and efficient protocol for the extraction of genomic DNA from plant pathogenic fungi was developed. Key features of the protocol include the SDS-assisted lysis of fungal mycelium with inclusion of a glass bead to help break hyphal walls, followed by isopropanol precipitation of the DNA. The protocol was used to extract genomic DNA from a collection of 26 fungal species, representing many important plant pathogens. Yield of DNA ranged from 2.1-4.9 g per 20 mg of mycelium or 0.4-0.6 g per 20 mg of spores. The DNA was of sufficient purity to be digested by restriction enzymes, to serve as a template in the PCR-amplification of genomic fragments as large as 4.9 kb, and to be used in dot-blot hybridization for the detection of multiple- and single-copy genes.
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