盐湖土壤微生物总DNA提取方法的比较  

Comparison of the modified metagenomic DNA extraction methods of microorganisms from salt lake soils

作　　者：苗莉云[1] 张鹏[2] 

机构地区：[1]运城学院生命科学系,山西运城044000 [2]湖北省农业科学院经济作物研究所,湖北武汉430064

出　　处：《浙江农业学报》2012年第6期1086-1090,共5页Acta Agriculturae Zhejiangensis

基　　金：国家自然科学基金(NSFC 20776058);中国博士后科学基金(20100471183);运城学院项目(20060025)

摘　　要：为了构建运城盐湖土壤微生物宏基因组文库,文章对盐湖土壤微生物总DNA的提取方法进行了比较和探索,通过改良6种植物DNA的提取方法,分别提取了盐湖土壤微生物总DNA,并采用紫外分光光度法、凝胶电泳、内切酶分析和PCR扩增法检测了各个方法所提取的DNA样品的质量。结果表明:改良后的方法 3是较适合提取运城盐湖土壤微生物总DNA的方法。采用该方法所得DNA的OD260/280值为1.883,DNA浓度和得率分别为18.1 ng·μL-1和72.4 ng·g-1;DNA片段长度约为23 kb,琼脂糖凝胶电泳条带清晰,无降解;DNA能被EcoRⅠ完全酶切,并可用于PCR扩增分析。In order to construct the metagenomic library of microorganisms from salt lake soils in Yuncheng, the mi- crobial metagenomic DNA extraction method was explored. Six of plant genomic DNA extraction methods were modi- fied to extract the microbial metagenomic DNA. The yield and quality of the metagenomie DNA was detected by ultra- violet spectrophotometry, gel electrophoresis, restriction endonuelease analysis and PCR amplification, and the re- sults showed that the modified method 3 was suitable for extracting the microbial metagenomic DNA from sah lake soils in Yuncheng. For the microbial metagenomic DNA extracted by the modified method 3, the value of 0D260/280 was 1. 883, the concentration and yield was 18.1 ng . μL-1 and 72.4 ng . g-1 respectively, and the length was about 23 kb with a clear stripe on the agarose gel. Moreover, the metagenomic DNA was completely digested with EcoR Ⅰ and also could be used for PCR amplification.

关 键 词：盐湖土壤 微生物 宏基因组 总DNA 提取方法 

分 类 号：S154.3[农业科学—土壤学]