4.1 Article

An inexpensive method for extraction of genomic DNA from fungal mycelia

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

  1. Alberta Canola Producers Commission
  2. Manitoba Canola Growers Association
  3. SaskCanola and the Canola Council of Canada
  4. Alberta Crop Industry Development Fund

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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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