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利用末端限制性片段长度多态性(T-RFLP)分析评估堆肥系统中的微生物群落结构。

Using terminal restriction fragment length polymorphism (T-RFLP) analysis to assess microbial community structure in compost systems.

作者信息

Tiquia Sonia M

机构信息

Department of Natural Sciences, University of Michigan, Dearborn, MI, USA.

出版信息

Methods Mol Biol. 2010;599:89-102. doi: 10.1007/978-1-60761-439-5_6.

Abstract

Terminal restriction fragment length polymorphism (T-RFLP) analysis of PCR-amplified genes is a widely used fingerprinting technique in composting systems. This analysis is based on the restriction endonuclease digestion of fluorescently end-labeled PCR products. The digested product is mixed with a DNA size standard, itself labeled with a distinct fluorescent dye, and the fragments are then separated by capillary or gel electrophoresis using an automated sequencer. Upon analysis, only the terminal end-labeled restriction fragments are detected. An electropherogram is produced, which shows a profile of compost microbial community as a series of peaks of varying height. This technique has also been effectively used in the exploration of complex microbial environments and in the study of bacterial, archaeal, and eukaryal populations in natural habitats.

摘要

对聚合酶链反应(PCR)扩增基因进行的末端限制性片段长度多态性(T-RFLP)分析是堆肥系统中广泛使用的指纹识别技术。该分析基于对荧光末端标记的PCR产物进行限制性内切酶消化。将消化后的产物与本身用不同荧光染料标记的DNA大小标准物混合,然后使用自动测序仪通过毛细管电泳或凝胶电泳分离片段。在分析时,仅检测末端标记的限制性片段。会生成一个电泳图,它将堆肥微生物群落的概况显示为一系列高度不同的峰。该技术也已有效地用于探索复杂的微生物环境以及研究自然栖息地中的细菌、古菌和真核生物种群。

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