| 盐湖土壤微生物总DNA提取方法的比较 |
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| 引用本文： | 苗莉云,张鹏.盐湖土壤微生物总DNA提取方法的比较[J].浙江农业学报,2012,24(6):0. |
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| 作者姓名： | 苗莉云 张鹏 |
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| 作者单位： | 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 |
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| 基金项目： | 国家自然科学基金，中国博士后科学基金，运城学院项目 |
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| 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。
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| 关 键 词： | 盐湖土壤 微生物 宏基因组 总DNA 提取方法 |
Comparison of the modified metagenomic DNA extraction methods of microorganisms from salt lake soils |
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| MIAO Li-yun
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ZHANG Peng.Comparison of the modified metagenomic DNA extraction methods of microorganisms from salt lake soils[J].Acta Agriculturae Zhejiangensis,2012,24(6):0. |
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| Authors: | MIAO Li-yun ZHANG Peng |
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| Abstract: | In order to construct the metagenomic library of microorganisms from salt lake soils in Yuncheng， the microbial metagenomic DNA extraction method was explored. Six of plant genomic DNA extraction methods were modified to extract the microbial metagenomic DNA. The yield and quality of the metagenomic DNA was detected by ultraviolet spectrophotometry， gel electrophoresis， restriction endonuclease analysis and PCR amplification， and the results showed that the modified method 3 was suitable for extracting the microbial metagenomic DNA from salt lake soils in Yuncheng. For the microbial metagenomic DNA extracted by the modified method 3， the value of OD260/280 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 EcoRⅠand also could be used for PCR amplification. |
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| Keywords: | salt lake soil microorganisms metagenome metagenomic DNA extraction method |
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