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Quantitative PCR for Detection of mRNA and gDNA in Environmental Isolates

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Environmental Microbiology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1096))

Abstract

Quantitative PCR is used to gauge the abundance of specific nucleic acid species within purified samples. Due to its high sensitivity and minimal operation costs, this method is routinely applied in modern molecular bioscience laboratories. Nonetheless, all quantitative PCR experiments must include several carefully designed, yet simple, controls to ensure the reliability of the analyses. The aim of this chapter is to provide basic quantitative PCR methods, from primer design through data analysis, that are generally applicable to studies in microbiology. These methods allow the abundance of targeted RNA or DNA molecules to be determined in nucleic acid samples purified from a variety of biological sources.

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Acknowledgements

Part of this work was supported by Project Grant 535053 from the National Health and Medical Research Council. KH is supported by an Australian Postdoctoral Fellowship funded by the Australian Research Council (grant DP110102680).

Author information

Authors and Affiliations

  1. School of Biological Sciences, University of Sydney, Sydney, NSW, Australia

    Anthony J. Brzoska

  2. Department of Chemistry and Biomolecular Sciences, Macquarie University, Sydney, NSW, Australia

    Karl A. Hassan

Authors
  1. Anthony J. Brzoska
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  2. Karl A. Hassan
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Editor information

Editors and Affiliations

  1. Dept. Chemistry & Biomolecular Sciences, Macquarie University, Sydney, New South Wales, Australia

    Ian T. Paulsen

  2. School of Molecular Bioscience, University of Sydney, Camperdown, New South Wales, Australia

    Andrew J. Holmes

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Brzoska, A.J., Hassan, K.A. (2014). Quantitative PCR for Detection of mRNA and gDNA in Environmental Isolates. In: Paulsen, I., Holmes, A. (eds) Environmental Microbiology. Methods in Molecular Biology, vol 1096. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-712-9_3

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  • DOI: https://doi.org/10.1007/978-1-62703-712-9_3

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-711-2

  • Online ISBN: 978-1-62703-712-9

  • eBook Packages: Springer Protocols

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