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Real-Time PCR – The Basic Principles

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Early, rapid and sensitive veterinary molecular diagnostics - real time PCR applications

Abstract

Conventional PCR is a powerful technique that allows exponential amplification of DNA sequences. A PCR reaction needs a pair of primers that are complementary to the sequence of interest. Primers are extended by the DNA polymerase . The copies produced after the extension, so called amplicons, are re-amplified with the same primers leading thus to an exponential amplification of the DNA molecules. After amplification, gel electrophoresis is used to analyse the amplified PCR products and this makes conventional PCR time consuming; since the reaction must finish before proceeding with the post-PCR analysis. Real Time PCR overcome this problem, because of its ability to measure the PCR amplicons at early states of the reaction as they are accumulate in a “Real Time Detection” mode thus measuring the amount of PCR product where the reaction is still in the exponential phase (QPCR).

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Authors and Affiliations

  1. Department of Nuclear Sciences and Applications, International Atomic Energy Agency (IAEA), A1400, Vienna, Austria

    Dr. Ericka A. Pestana, Dr. Adama Diallo, Dr. John R. Crowther & Prof. Gerrit J. Viljoen

  2. Department of Virology, Swedish University of Agricultural Sciences, SE-750 07, Uppsala, Sweden

    Prof. Sandor Belak

Authors
  1. Dr. Ericka A. Pestana
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  2. Prof. Sandor Belak
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  3. Dr. Adama Diallo
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  4. Dr. John R. Crowther
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  5. Prof. Gerrit J. Viljoen
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Correspondence to Ericka A. Pestana .

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Pestana, E.A., Belak, S., Diallo, A., Crowther, J.R., Viljoen, G.J. (2009). Real-Time PCR – The Basic Principles. In: Early, rapid and sensitive veterinary molecular diagnostics - real time PCR applications. Springer, Dordrecht. https://doi.org/10.1007/978-90-481-3132-7_3

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