摘要
[目的]对土壤微生物的DNA进行提取,以建立一种适用于PCR技术分析的土壤微生物DNA的快速提取方法。[方法]采用了2种不同的直接裂解法提取土壤微生物DNA,并用细菌通用引物27F和1492R进行PCR扩增16S rDNA片段。[结果]2种方法提取的土壤微生物总DNA电泳结果显示没有差别,但是只有CTAB、溶菌酶、冻融裂解法提取的总DNA不需要纯化,在增加Mg2+用量的条件下,只需用第一轮非特异PCR产物作为模板即可扩增出16S rDNA片段。[结论]CTAB、溶菌酶、冻融裂解法提取土壤DNA,方法操作简单、快捷,同时适用于PCR分析,为土壤微生物群落结构的多样性分析奠定了基础。
[ Objective ] The aim of this study was to extract DNA from soil microorganisms and establish the rapid extraction method of soil microbial DNA for PCR analysis. [ Method] Two kinds of different direct pyrolysis methods were investigated. 16S rDNA was amplified with bacterial universal primers 27F and 1492R by PCR. [ Result] The electrophoresis results of total DNA from soil microorganisms by two kinds of DNA extraction methods had no difference. Without purification of Total DNA extracted by CTAB, lysozyme, freezing thaw pyrolysis method,the first-cycle non-specific PCR products could be used as the template to amplify 16S rDNA fragments under the conditions of adding Mg^2＋ amount. [ Conclusion ] Soil DNA could extracted by CTAB ,lysozyme ,freezing thaw pyrolysis method. The method was simple and fast in operation,suitable for PCR analysis. The research provided the basis for the diversity analysis of soil microbial communities.
出处
《安徽农业科学》
CAS
北大核心
2010年第2期600-601,619,共3页
Journal of Anhui Agricultural Sciences
基金
广西师范大学第三批青年骨干教师资助计划(2007)
