Advantages and limitations of quantitative PCR (Q-PCR)-based approaches in microbial ecology

Smith, Cindy J. and Osborn, A. Mark (2009) Advantages and limitations of quantitative PCR (Q-PCR)-based approaches in microbial ecology. FEMS Microbiology Ecology, 67 (1). pp. 6-20. ISSN 0168-6496

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Item Type:Article
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Abstract

Quantitative PCR (Q-PCR or real-time PCR) approaches are now widely applied in microbial ecology to quantify the abundance and expression of taxonomic and functional gene markers within the environment. Q-PCR-based analyses combine 'traditional' end-point detection PCR with fluorescent detection technologies to record the accumulation of amplicons in 'real time' during each cycle of the PCR amplification. By detection of amplicons during the early exponential phase of the PCR, this enables the quantification of gene (or transcript) numbers when these are proportional to the starting template concentration. When Q-PCR is coupled with a preceding reverse transcription reaction, it can be used to quantify gene expression (RT-Q-PCR). This review firstly addresses the theoretical and practical implementation of Q-PCR and RT-Q-PCR protocols in microbial ecology, highlighting key experimental considerations. Secondly, we review the applications of (RT)-Q-PCR analyses in environmental microbiology and evaluate the contribution and advances gained from such approaches. Finally, we conclude by offering future perspectives on the application of (RT)-Q-PCR in furthering understanding in microbial ecology, in particular, when coupled with other molecular approaches and more traditional investigations of environmental systems. © 2008 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

Additional Information:Article first published online: 9 DEC 2008
Keywords:gene expression, genetic marker, microbial ecology, polymerase chain reaction, RNA, amplicon, fluorescence analysis, gene amplification, genetic transcription, nonhuman, priority journal, quantitative analysis, real time polymerase chain reaction, short survey, Ecology, Environmental Microbiology, Reverse Transcriptase Polymerase Chain Reaction, RNA, Messenger, RNA, Ribosomal, 16S
Subjects:C Biological Sciences > C180 Ecology
C Biological Sciences > C500 Microbiology
Divisions:College of Science > School of Life Sciences
ID Code:8948
Deposited On:22 Apr 2013 09:13

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