| 活性污泥基因组DNA快速提取新方法及其指纹分析 |
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| 引用本文： | 曲媛媛,周集体,王竞,夏元化,柳广飞,谭靓.活性污泥基因组DNA快速提取新方法及其指纹分析[J].大连理工大学学报,2006,46(3):335-339. |
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| 作者姓名： | 曲媛媛 周集体 王竞 夏元化 柳广飞 谭靓 |
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| 作者单位： | 大连理工大学,环境与生命学院,辽宁,大连,116024 |
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| 摘 要： | 建立了从活性污泥中快速提取基因组DNA的新方法，过程包括粗提与精制两个步骤，简化了繁琐的提取程序，提高了提取效率．用该方法提取的基因组DNA做模板，进行扩增核糖体限制性酶切片断分析（ARDRA）及核糖体基因间区序列分析（RISA），均获得了理想的多态性指纹图谱；采用两对特异性引物（鞘氨醇单胞菌属和氨加氧酶基因）对所提污泥基因组DNA进行扩增，均获得了正确的PCR产物．该方法提取的污泥基因组DNA不但适用于研究污泥系统中菌群多样性分析，而且还可以对特定基因进行跟踪监测。
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| 关 键 词： | 活性污泥 基因组DNA提取 |
| 文章编号： | 1000-8608（2006）02-0335-05 |
| 收稿时间： | 2005-01-25 |
| 修稿时间： | 2005-01-252006-02-21 |
Novel approach for rapid extraction of genomic DNA from activated sludge and its fingerprinting analysis |
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| QU Yuan-yuan,ZHOU Ji-ti,WANG Jing,XIA Yuan-hua,LIU Guang-fei,TAN Liang.Novel approach for rapid extraction of genomic DNA from activated sludge and its fingerprinting analysis[J].Journal of Dalian University of Technology,2006,46(3):335-339. |
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| Authors: | QU Yuan-yuan ZHOU Ji-ti WANG Jing XIA Yuan-hua LIU Guang-fei TAN Liang |
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| Institution: | School of Environ. and Biol. Sci. and Technol., Dalian Univ. of Technol., Dalian 116024, China |
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| Abstract: | A novel approach for rapid extraction of genomic DNA from activated sludge was developed.The extraction protocol comprised two steps: crude extraction and further purification,which simplified the complicated extraction process.The satisfied multiple fingerprints were obtained by this template DNA for both ARDRA(amplified ribosomal DNA restriction analysis) and RISA(ribosomal intergenic space analysis).Simultaneously,two pairs of specific primers(Sphingomonas sp. and amo gene) were used to perform PCR,and the correct PCR products were obtained.The proposed protocol was rapid and efficient not only for analysis of microbial community diversity,but also for monitoring specific gene in activated sludge. |
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| Keywords: | ARDRA RISA |
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