| PCR-DGGE应用于土壤生态学研究的流程及优化 |
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| 引用本文： | 许文欢,阮宏华.PCR-DGGE应用于土壤生态学研究的流程及优化[J].江苏林业科技,2014(4):46-49. |
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| 作者姓名： | 许文欢 阮宏华 |
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| 作者单位： | 南京林业大学林学院;南京林业大学江苏省林业生态工程重点实验室; |
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| 基金项目： | 国家自然科学基金项目(31170417);江苏省研究生培养创新工程项目(CXZZ_110510);江苏优势学科建设和现代林业协同创新项目;江苏省自然科学基金青年专项项目(BK20130974) |
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| 摘 要： | 土壤微生物的多样性在生态学研究中越来越受到关注。能反映微生物多样性的技术有很多,PCR-变性梯度凝胶电泳(Polymerase chain reaction-denaturing gradient gel electrophoresis,PCR-DGGE)技术是其中的一种。此技术有其独到特色,广泛应用于各个领域。目前关于该技术的应用已有许多报道,然而缺少对该技术应用于土壤生态学研究详细操作流程的归纳和总结。操作过程中有许多技术性问题,包括如何提取和纯化土壤DNA、PCR反应条件设置、DGGE电泳条件设置以及图谱数据分析等。该文简要归纳了PCR-DGGE技术应用于土壤生态学研究的流程以及如何优化关键性步骤,并提出了有待优化的方向。
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| 关 键 词： | PCR-DGGE 土壤微生物 生态学 流程 优化 |
Procedures and optimization of PCR-DGGE applying to the research of soil ecology |
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| Institution: | XU Wen-huan, RUAN Hong-hua ( 1. College of Forestry, Nanjing Forestry University, Nanjing 210037, China; 2. Key Laboratory of Forestry Ecological Engineering of Jiangsu Province, Nanjing Forestry University, Nanjing 210037, China) |
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| Abstract: | Soil microbial diversity has received more and more attention in the research of ecology. As one of the techniques which can be applied to present the diversity of microorganism,PCR-denaturing gradient gel electrophoresis（PCR-DGGE）has its own feature and has been widely used in various fields. So far,there were so many reports on the use of this technology but few ones on the summary of detailed operation procedures. During the operation procedures,there were some technical problems including how to extract and purify soil DNA,how to set the parameters of the conditions of PCR,how to set the condition of DGGE electrophoresis,how to analyze the atlas and data and so on. The procedures of PCR-DGGE applying to the research of soil ecology,as well as method to optimize critical procedures and the direction of optimization,were reviewed in this paper. |
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| Keywords: | PCR-DGGE Soil microorganism Ecology Procedure Optimization |
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