A simple method for a mini-preparation of fungal DNA |
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Authors: | Ken-ichiro Saitoh Kana Togashi Tsutomu Arie Tohru Teraoka |
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Affiliation: | (1) Graduate School of Agriculture, Tokyo University of Agriculture and Technology, Tokyo, Japan;(2) Laboratory of Plant Pathology, Faculty of Agriculture, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu, Tokyo 183-8509, Japan |
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Abstract: | A simple method was established to prepare DNA from fungal mycelia cultured on an agar plate. The fungi tested successfully with this method contained Zygomycetes, Ascomycetes, Basidiomycetes, and Oomycetes. This method did not require any time-consuming steps to crush or digest mycelia or fractionation in a phenol–chloroform mixture. The DNA was easily extracted by immersing and dispersing the mycelial plugs in a specific buffer (200 mM Tris-HCl, 50 mM ethylenediaminetetraacetic acid, 200 mM NaCl, 1% n-lauroylsarcosine, pH 8.0), then concentrated by ethanol precipitation. The total time to complete the whole procedure was less than 1 h. The quality and quantity were sufficient for polymerase chain reaction amplification and Southern blot analysis. |
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Keywords: | Fungal DNA extraction Zygomycetes Ascomycetes Basidiomycetes Oomycetes |
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