| 厌氧活性污泥PCR-DGGE图谱的优化 |
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| 引用本文： | 魏利,马放,刘亚丽,杨远彪,王丽娜,翟旭.厌氧活性污泥PCR-DGGE图谱的优化[J].哈尔滨工业大学学报,2009,41(2):81-84. |
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| 作者姓名： | 魏利 马放 刘亚丽 杨远彪 王丽娜 翟旭 |
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| 作者单位： | 魏利,马放,翟旭,WEI Li,MA Fang,ZHAI Xu(哈尔滨工业大学,市政环境工程学院,哈尔滨,150090);刘亚丽,王丽娜,LIU Ya-li,WANG Li-na(东北林业大学,林学院,哈尔滨,150040);杨远彪,YANG Yuan-biao(广西大学,林学院,南宁,530004)
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| 基金项目： | 国家重点基础研究发展规划(973计划) |
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| 摘 要： | 针对特种污泥PCR-DGGE图谱的优化一直是微生物分子生态学研究的重点.在反硝化抑制硫酸盐还原菌的研究中,研究厌氧硫酸盐还原污泥和反硝化污泥DNA的提取方法,应用通用引物958F/1401R扩增16S rDNA序列,探讨不同的退火温度对DGGE指纹图谱多样性影响,同时采用时间进程法对DGGE图谱进行优化.结果表明,采用PBS洗涤污泥和超声震荡有利于硫酸盐还原污泥DNA提取;采用958F/1401R为引物时,扩增片段大约500 bp,不同的退火温度对DGGE指纹图谱的多样性影响不显著;采用时间进程法可以很好地再现不同电泳阶段对样品的分离效果,最佳变性剂梯度为40%～60%,在130 V电压下,最佳电泳时间为5 h.
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| 关 键 词： | DGGE 16S rDNA 退火温度 时间进程法 活性污泥 |
Optimization of profiles of PCR-denaturing gradient gel electrophoresis for anaerobic activated sludge |
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| WEI Li,MA Fang,LIU Ya-li,YANG Yuan-biao,WANG Li-na,ZHAI Xu.Optimization of profiles of PCR-denaturing gradient gel electrophoresis for anaerobic activated sludge[J].Journal of Harbin Institute of Technology,2009,41(2):81-84. |
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| Authors: | WEI Li MA Fang LIU Ya-li YANG Yuan-biao WANG Li-na ZHAI Xu |
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| Affiliation: | 1(1.School of Municipal and Environmental Engineering,Harbin Institute of Technology,Harbin 150090,China;2.School of Forestry,Northeast Forestry University,Harbin 150040,China;3.School of Forestry,Guangxi University,Nanning 530004,China) |
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| Abstract: | The optimization of PCR-DGGE atlas for special sludge has attracted the emphasis of the investigation of microbial molecular ecology.In the research of denitrification inhibiting sulfate-reducing bacteria,DNA extracting method of anaerobic sulfate reducing sludge and denitrifying sludge was studied.By amplifying 16S rDNA sequence with universal primer 958F/1401R,the influences of different annealing temperatures on DGGE fingerprint were investigated,and the DGGE atlas was optimized by time travel.The results indicate that it is beneficial to the DNA extraction from sulfate reducing sludge by PBS washing sludge and sonic oscillation;the amplified segment is about 500 bp with the primer 958F/1401R,and the influences of different annealing temperatures on DGGE fingerprint are not obvious;the isolating effects of different electrophoresis stages for samples can be reappeared very well by time travel,and the best denaturing gradient ranks from 40% to 60%,the best electrophoresis time is 5 h under 130 V voltage. |
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| Keywords: | DGGE 16SrDNA |
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