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How Many Microorganisms Are Present? Quantitative Reverse Transcription PCR (qRT-PCR)

Price, A and �lvarez, LA and Whitby, C and Larsen, J (2010) 'How Many Microorganisms Are Present? Quantitative Reverse Transcription PCR (qRT-PCR).' In: Whitby, C and Skovhus, TL, (eds.) Applied Microbiology and Molecular Biology in Oilfield Systems. Springer, 77 - 84. ISBN 978-90-481-9251-9

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Abstract

Quantitative reverse transcription PCR (qRT-PCR) is a variation of conventional quantitative or real-time PCR, whereby mRNA is first converted into the complementary DNA (cDNA) by reverse transcription, the cDNA is then subsequently quantified by qPCR. The use of mRNA as the initial template allows the quantification of gene transcripts, rather than gene copy numbers. mRNA is only produced by actively metabolising cells and is produced by its corresponding gene to provide a ?blueprint? in order for a cell to manufacture a specific protein. Conventional qPCR detects not only DNA present in actively metabolising cells but also inactive and dead cells. qRT-PCR has the advantage that only actively metabolising cells are detected, hence provides a more reliable measure of microbial activity in oilfield samples. When qRT-PCR is combined with primers and probes for specific genes, the activity of microbial processes important in the oilfield, such as sulphate reduction, methanogenesis and nitrate reduction can be monitored.

Item Type: Book Section
Additional Information: Proceedings from the International Symposium on Applied Microbiology and Molecular Biology in Oil Systems (ISMOS-2), 2009
Subjects: Q Science > QH Natural history > QH301 Biology
Divisions: Faculty of Science and Health > Biological Sciences, School of
Depositing User: Jim Jamieson
Date Deposited: 13 Jan 2016 11:49
Last Modified: 17 Aug 2017 17:29
URI: http://repository.essex.ac.uk/id/eprint/15838

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