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

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.