农田土壤中病原真菌DNA提取方法的研究
Study on an extraction method of pathogenic fungal DNA from farmland soil
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摘要: 通过比较不同的研磨、洗涤、裂解和沉淀方法,成功获得一种适于土壤真菌DNA提取的方法.结果表明,采用石英砂研磨、TENP和PBS联合洗涤、SDS高盐裂解、异丙醇沉淀等一系列步骤,所获取的土壤微生物DNA质量最佳.该试验方法可以直接从土壤中提取微生物基因组,尤其适用于土壤病原真菌的提取,所得DNA完全可以直接用于酶切和PCR扩增.该方法高效、价廉、操作简单,对改进土壤病原菌生态的研究方法具有重要价值.Abstract: By comparing grinding method, washing method, cleavage and precipitation method, a method was gotten suitable for farmland fungi DNA extraction. The results showed that the quality of soil microorganism DNA is of the best which is obtained by grinding with quartz sand, washing with TENP and PBS, lysing with SDS in high-salt buffer and precipitating with isopropanol and so on. The pathogenic fungal DNA can be directly extracted from soil by using the method, which is particularly suitable for the soil pathogenic fungal extraction. The DNA obtained can be directly used for restriction enzyme digestion and PCR amplification. The method is efficient, inexpensive, easy to operate, which is of important value for the soil pathogens ecological research.
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Key words:
- farmland soil /
- pathogenic fungi /
- DNA extraction /
- PCR amplification
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