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Medical Biomethods Handbook pp 305–313Cite as

Quantitative TaqMan Real-Time PCR

Diagnostic and Scientific Applications

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Abstract

The invention of real-time polymerase chair reaction (PCR) has revolutionized the quantification of gene expression and DNA copy number measurements. However, after the first documentation of real-time PCR in 1993 (1), it took several years for this method to become a mainstream tool. PCR generates DNA copies in an exponential way. As soon as resources are exhausted, however, the so-called plateau phase of PCR reaction is reached, making quantification very unreliable. Therefore, quantification appears most reliable in the early exponential phase of PCR (i.e., in a “real-time” fashion). To ensure measurements in this phase of the PCR cycle, real-time PCR measures as soon as the threshold of detection is definitely reached. The cycle of PCR at which this occurs is then named the threshold cycle (2) (see Fig. 1). It is the objective of this chapter to describe the possibilities of TaqMan real-time PCR for mRNA and DNA quantification and to discuss pitfalls and alternatives.

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Author information

Authors and Affiliations

  1. Department of Pediatrics, University of Erlangen, Erlangen, Germany

    Jörg Dötsch, Ellen Schoof & Wolfgang Rascher

Authors
  1. Jörg Dötsch
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  2. Ellen Schoof
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  3. Wolfgang Rascher
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Editor information

Editors and Affiliations

  1. University of Hertfordshire, Hatfield, UK

    John M. Walker  & Ralph Rapley  & 

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© 2005 Humana Press Inc., Totowa, NJ

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Dötsch, J., Schoof, E., Rascher, W. (2005). Quantitative TaqMan Real-Time PCR. In: Walker, J.M., Rapley, R. (eds) Medical Biomethods Handbook. Springer Protocols Handbooks. Humana Press. https://doi.org/10.1385/1-59259-870-6:305

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  • DOI: https://doi.org/10.1385/1-59259-870-6:305

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-288-9

  • Online ISBN: 978-1-59259-870-0

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