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Extraction and PCR Amplification of DNA from Minute Ectoparasitic Fungi

Alex Weir and Meredith Blackwell
Mycologia
Vol. 93, No. 4 (Jul. - Aug., 2001), pp. 802-806
DOI: 10.2307/3761835
Stable URL: http://www.jstor.org/stable/3761835
Page Count: 5
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Since scans are not currently available to screen readers, please contact JSTOR User Support for access. We'll provide a PDF copy for your screen reader.
Extraction and PCR Amplification of DNA from Minute Ectoparasitic Fungi
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Abstract

Laboulbeniales is a large order of ascomycete fungi characterized by microscopic size, determinate growth patterns, thalloid morphology, and obligately parasitic lifestyle that complicate the application of molecular phylogenetic methods. Here, we describe improved techniques for the isolation of genomic DNA from Laboulbeniales, and use of the resulting extracts for PCR amplification of 18S rDNA and 28S rDNA. The important modification we report is the extraction and retrieval of DNA on dry ice; the dramatic improvements in consistency and yield indicate that these protocols could be applied successfully to a wide range of small, melanized organisms that cannot be grown in culture.

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