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

Smith, C and Osborn, A 2009, 'Advantages and limitations of quantitative PCR (Q-PCR)-based approaches in microbial ecology', FEMS Microbiology Ecology, vol. 67, no. 1, pp. 6-20.


Document type: Journal Article
Collection: Journal Articles

Title Advantages and limitations of quantitative PCR (Q-PCR)-based approaches in microbial ecology
Author(s) Smith, C
Osborn, A
Year 2009
Journal name FEMS Microbiology Ecology
Volume number 67
Issue number 1
Start page 6
End page 20
Total pages 15
Publisher Wiley-Blackwell Publishing
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
Subject Biological Sciences not elsewhere classified
Keyword(s) 16S rRNA gene
mRNA
Q-PCR
RT-Q-PCR
DOI - identifier 10.1111/j.1574-6941.2008.00629.x
Copyright notice © 2008 Federation of European Microbiological Societies
ISSN 0168-6496
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