一种适于PCR反应的快速提取食用菌基因组DNA的方法

A Rapid Method for Extraction of Genomic DNA of Mushrooms for PCR Analysis

目的:建立一种以食用菌菌丝体和子实体为原材料的快速提取其基因组DNA的方法,从而提高基因组DNA的提取效率,为食用菌分子生物学提供便利。方法:分别以平菇黑平王的菌丝体和子实体为材料,快速提取其基因组DNA,并以此为模板进行ITS序列扩增。结果:采用方法提取的基因组DNA结构完整,无明显拖尾现象,浓度大约为10ng/μl,以此为模板能够获得预期的ITS条带。结论:该方法具有简便、快速、经济、无污染等优点,提取的基因组DNA适用于PCR反应等分子生物学研究,提高了提取效率,可作为高通量的提取方法。

Objective：To extablish a rapid method to extract the mycelium or fruiting body of mushroom.Thus,this method can accelerate the efficiency of the extraction and be well served for the molecular biology of the mushrooms.Method：Genomic DNA were extracted from mycelium and fruiting body of Pleurotus ostreatus and assayed by amplification of ITS sequence using these genomic DNA as template.Result：The extracted DNA was structural integrity and the concentration was about 10 ng/μl.The genomic DNA extracted by using this method could be sufficiently for ITS amplification.Conclusion：The results showed that it was a powerful,rapid,economically,non-pollution method for the genome DNA＇s extraction for mycelium and fruiting body of Pleurotus ostreatus.The results indicated that this method was an efficient method for the isolation of the genomic DNA of mushroom.