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Polymerase Chain Reaction

  • Chapter
Techniques in Molecular Medicine

Part of the book series: Springer Lab Manual ((SLM))

Abstract

The polymerase chain reaction (PCR) (1) is a rapid technique for in vitro amplification of a specific DNA fragment by use of two short single- stranded primers flanking this fragment. Through repeated cycles of heat denaturation of the double-stranded DNA template, primer annealing, and primer extension using a heat-stable polymerase, the fragment of interest is amplified exponentially up to a million fold. Starting from very small amounts of DNA such as even that contained in a single cell, µg amounts of PCR product may be produced. The PCR product can be further modified by cloning into a vector, labeling for use as a probe, or directly sequenced. Starting with very small amounts of template the technique gives a high yield and can be modified in many ways. In this chapter the components and mechanisms of a PCR are described, followed by a standard PCR proctocol. Subsequently, modifications of this protocol for specific experimental purposes are discussed.

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Authors and Affiliations

  1. Universitäts-Kinderklinik, Mathildenstrasse 1, Freiburg, 79106, Germany

    Friedhelm Hildebrandt

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  1. Friedhelm Hildebrandt
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  2. Iva Singh-Sawhney
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Correspondence to Friedhelm Hildebrandt .

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Editors and Affiliations

  1. Universitäts-Kinderklinik, Mathildenstrasse 1, D-79106, Freiburg, Germany

    Friedhelm Hildebrandt

  2. Section of Nephrology, Dep. of Internal Medicine, Yale University, 2073 Laboratory of Medicine and Pediatrics, 333 Cedar Street, New Haven, Connecticut, 06620-8029, USA

    Peter Igarashi M.D.

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© 1999 Springer-Verlag Berlin Heidelberg

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Hildebrandt, F., Singh-Sawhney, I. (1999). Polymerase Chain Reaction. In: Hildebrandt, F., Igarashi, P. (eds) Techniques in Molecular Medicine. Springer Lab Manual. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-59811-1_14

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  • DOI: https://doi.org/10.1007/978-3-642-59811-1_14

  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-642-47808-6

  • Online ISBN: 978-3-642-59811-1

  • eBook Packages: Springer Book Archive

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