Skip to main content
SpringerLink
Log in

Rapid preparation of PCR-amplifiable fungal DNA by benzyl bromide extraction

  • Note
  • Published:
Mycoscience

Abstract

For isolation of fungal DNA for PCR amplification, we compared three DNA isolation methods: enzymatic cleavage and the use of benzyl chloride or benzyl bromide. Since benzyl bromide is more reactive, its use enabled us to readily isolate the total nucleic acids as a DNA template source from various fungi, including dematiaceous hyphomycetes, for RAPD analysis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Literature cited

  • Biel, S. W. and Parrish, F. W. 1986. Isolation of DNA from fungal mycelia and sclerotia without use of density gradient ultracentrifugation. Anal. Biochem.154: 21–25.

    Article  PubMed  Google Scholar 

  • Fujimori, F. and Okuda, T. 1994. Application of the random amplified polymorphic DNA using the polymerase chain reaction for efficient elimination of duplicate strains in microbial screening. I. Fungi. J. Antibiot.47: 173–182.

    PubMed  Google Scholar 

  • Goodwin, P. H. and Annis, S. L. 1991. Rapid identification of genetic variation and pathotype ofLeptosphaeria maculans by random amplified polymorphic DNA assay. Appl. Environ. Microbiol.57: 2482–2486.

    PubMed  Google Scholar 

  • Hudspeth, M. E. S., Shumard, D. S., Tatti, K. M. and Grossman, L. I. 1980. Rapid purification of yeast mitochondrial DNA in high yield. Biochim. Biophys. Acta610: 221–228.

    PubMed  Google Scholar 

  • Lehmann, P. F., Lin, D. and Lasker, B. A. 1992. Genotypic identification and characterization of species and strains within the genusCandida by using random amplified polymorphic DNA. J. Clin. Microbiol.30: 3249–3254.

    PubMed  Google Scholar 

  • Okuda, T., Yanagisawa, M., Fujimori, F., Nishizuka, Y., Takehana, Y. and Sugiyama, M. 1995. New isolation methods and PCR strain identification techniques for natural products screening programs. Can. J. Bot. (In press.)

  • Omori, Y., Tsumura, M., Ichida, T. and Yamasato, K. 1990. Disrupting of fungus and actinomycetes cells with Cryo Clean Blaster (CCB). Jpn. J. Freez. Dry.36: 36–40.

    Google Scholar 

  • Welsh, J., Peterson, C. and McClelland, M. 1991. Polymorphisms generated by arbitrarily primed PCR in the mouse: application to strain identification and genetic mapping. Nucleic Acids Res.19: 303–306.

    PubMed  Google Scholar 

  • Williams, J. G., Kubelik, K. A. R., Livak, K. J., Rafalski, J. A. and Tingey, S. V. 1990. DNA polymorphisms amplified by arbitrary primer are useful as genetic markers. Nucleic Acids Res.18: 6531–6535.

    PubMed  Google Scholar 

  • Yoon, C-S., Glawe, D. A. and Shaw, P. D. 1991. A method for rapid small-scale preparation of fungal DNA. Mycologia83:835–838.

    Google Scholar 

  • Zhu, H., Qu, F. and Zhu, L-H. 1993. Isolation of genomic DNAs from plants, fungi and bacteria using benzyl chloride. Nucleic Acids Res.21: 5279–5280.

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Microbiology and Taxonomy, Nippon Roche Research Center, 200, Kajiwara, 247, Kamakura, Kanagawa, Japan

    Fumihiro Fujimori & Toru Okuda

Authors
  1. Fumihiro Fujimori
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Toru Okuda
    View author publications

    You can also search for this author in PubMed Google Scholar

About this article

Cite this article

Fujimori, F., Okuda, T. Rapid preparation of PCR-amplifiable fungal DNA by benzyl bromide extraction. Mycoscience 36, 465–467 (1995). https://doi.org/10.1007/BF02268634

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02268634

Key Words

Search

Navigation