A simple method for a mini-preparation of fungal DNA
A simple method for a mini-preparation of fungal DNA
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DOI：
10.1007/s10327-006-0300-1
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发表时间：
2006-12-01
影响因子：
1.2
通讯作者：
Teraoka, Tohru
中科院分区：
文献类型：
--
作者：
Saitoh, Ken-ichiro;Togashi, Kana;Teraoka, Tohru
建立了一种从琼脂平板上培养的真菌菌丝制备DNA的简便方法。用这种方法成功测试的真菌包括接合菌、子囊菌、担子菌和卵菌。这种方法不需要任何耗时的步骤来粉碎或消化菌丝体或在苯酚-氯仿混合物中分离。将菌丝塞浸泡并分散在特定的缓冲液（200mM Tris-HCl， 50mM乙二胺四乙酸，200mM NaCl， 1% n-月桂酰肌氨酸，pH 8.0）中，然后用乙醇沉淀浓缩。完成整个过程的总时间不到1小时。质量和数量足以进行聚合酶链反应扩增和Southern blot分析。
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 (200mM Tris-HCl, 50mM ethylenediaminetetraacetic acid, 200mM NaCl, 1% n-lauroylsarcosine, pH 8.0), then concentrated by ethanol precipitation. The total time to complete the whole procedure was less than 1h. The quality and quantity were sufficient for polymerase chain reaction amplification and Southern blot analysis.