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Overview of Limitation and Improving Methods of PCR-DGGE in Microbial Diversity Study

Jian Kong

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Abstract

Polymerase chain reaction-denaturing gradient gel electrophoresis(PCR-DGGE) technique has been proven to be a powerful tool for analyzing microbial communities and their dynamics changes.As a fast and reliable molecular method,DGGE is used widely to investigate the microbial component and structure of complex communities without isolation and cultivation processes.However,potential causes resulting in limitations in DGGE analysis can be sometimes met.In this paper,the principle and application aspects,as well as the limitation of DGGE techniques for microbial ecology analysis were introduced,and the main causing factors addressed and discussed,such as DNA extraction,PCR amplification,DGGE conditions optimization,and DGGE profile analysis.

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What this paper is about

Polymerase chain reaction-denaturing gradient gel electrophoresis(PCR-DGGE) technique has been proven to be a powerful tool for analyzing microbial communities and their dynamics changes.As a fast and reliable molecular method,DGGE is used widely to investigate the microbial component and structure of complex communities without isolation and cultivation processes.However,potential causes resulting in limitations in DGGE analysis can be sometimes met.In this paper,the principle and application aspects,as well as the limitation of DGGE techniques for microbial ecology analysis were introduced,and the main causing factors addressed and discussed,such as DNA extraction,PCR amplification,DGGE conditions optimization,and DGGE profile analysis.

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Available abstract

Polymerase chain reaction-denaturing gradient gel electrophoresis(PCR-DGGE) technique has been proven to be a powerful tool for analyzing microbial communities and their dynamics changes.As a fast and reliable molecular method,DGGE is used widely to investigate the microbial component and structure of complex communities without isolation and cultivation processes.However,potential causes resulting in limitations in DGGE analysis can be sometimes met.In this paper,the principle and application aspects,as well as the limitation of DGGE techniques for microbial ecology analysis were introduced,and the main causing factors addressed and discussed,such as DNA extraction,PCR amplification,DGGE conditions optimization,and DGGE profile analysis.

Key concepts: Temperature gradient gel electrophoresis, Isolation (microbiology), Microbial ecology, Biology, Microbial population biology, Polymerase chain reaction, Biochemical engineering, Microbiology

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