西北地区耕地土壤真菌DNA提取方法比较  

Comparison of DNA Extraction Methods for Soil Fungal DNA from Arable Land Soils in Northwest China

作　　者：汤世博[1] 张仁陟[1] 张金文[2] 

机构地区：[1]甘肃农业大学资源与环境学院,甘肃兰州730070 [2]甘肃农业大学农学院,甘肃兰州730070

出　　处：《土壤通报》2012年第4期832-835,共4页Chinese Journal of Soil Science

基　　金：ACIAR资助项目(SMCN(LWR2)/1999/094);甘肃省农牧厅资助项目(034048);中国农村技术开发中心资助项目(0390993)资助

摘　　要：由于西北土壤理化性质的复杂性和真菌特殊性,所以从土壤中提取真菌基因组DNA就相对细菌更困难。在2种常用的土壤微生物基因组DNA提取方法与在传统提取方法的基础上,结合了一种专门适用于真菌的提取方法进行了比较,并且利用真菌28SrDNA通用引物U1/U2进行扩增。三种提取方法比较结果表明:SDS法提取的DNA纯度最低,传统CTAB-SDS的DNA产量最低,实验室的提取方法既可以提高DNA产量又可以保证DNA的片段完整性,并且本实验室的提取方法扩增效果最好,可广泛应用于西北地区土壤真菌的分子生物学研究。Owing to the complexity of Northwest soil physical and chemical properties and the particularity of fungi, the extraction of fungi DNA genome is more difficult than that of bacterial from soil samples. In this paper, the two useful kinds of soil microbial genomic DNA extraction methods and the combination of a specialized extraction method for fungi based on traditional method were compared, and using fungal universal primers U1/U2 28SrDNA amplification. The results of comparison of three different methods showed that DNA extracted by SDS method obtained the lowest purity, and by the traditional CTAB-SDS method provided the lowest DNA yield. The extraction of the laboratory method could enhance the DNA extraction yield and guarantee the integrity of DNA fragments, and obtained the best effect for PCR amplification and be widely used in fungi molecular biology of Northwest soils.

关 键 词：真菌 西北土壤 基因组DNA 

分 类 号：S154.3[农业科学—土壤学]