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High Resolution PCR Quantitation by AmpliSensor Assay

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Quantitative PCR Protocols

Part of the book series: Methods in Molecular Medicine™ ((MIMM,volume 26))

Abstract

A real-time kinetic tool for polymerase chain reaction (PCR) quantitation, AmpliSensor assay (1) quantifies PCR product by relating the rate of an amplification reaction through the progressive depletion of a rate-limiting primer. AmpliSensor assay invokes a two-step amplification scheme:

  1. 1.

    An initial amplification with one primer in excess in order to overproduce one strand of the target, and

  2. 2.

    A subsequent hemi-nested amplification for fluorescence signal detection.

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References

  1. Wang, C. N., Wu, K. Y., and Wang, H.-T. (1995) Quantitative PCR using the AmpliSensor assay, in PCR Primer: A Laboratory Manual (Diefennbach, C. W. and Dveksler, G. S., eds.), Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, pp. 193–202.

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  4. Chiang, P.-W., Song, W. L., Wu, K. Y., Korenberg, J. R., Fogel, E. J., Van Keuren, H. L., Lashkari, D., and Kurnit, D. M. (1996) Use of a fluorescence-PCR reaction to detect genomic sequence copy number and transcriptional abundance. Genome Methods 6, 1013–1026.

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  5. Chen, S., Yee, A., Griffiths, M., Wu, K. Y., Wang, C. N., Rahn, K., and De Grandis, S. A. (1997) A rapid, sensitive and automated method for detection of salmonella species in foods using AG-9600 AmpliSensor analyzer. J. Appl. Microbiol. 83, 314–321.

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Author information

Authors and Affiliations

  1. Biotronics Corporation, Lowell, MA

    Chang-Ning Wang

Authors
  1. Chang-Ning Wang
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Editor information

Editors and Affiliations

  1. University of Regensburg, Germany

    Bernd Kochanowski  & Udo Reischl  & 

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© 1999 Humana Press Inc., Totowa, NJ

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Wang, CN. (1999). High Resolution PCR Quantitation by AmpliSensor Assay. In: Kochanowski, B., Reischl, U. (eds) Quantitative PCR Protocols. Methods in Molecular Medicine™, vol 26. Humana Press. https://doi.org/10.1385/0-89603-518-2:245

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  • DOI: https://doi.org/10.1385/0-89603-518-2:245

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-518-8

  • Online ISBN: 978-1-59259-262-3

  • eBook Packages: Springer Protocols

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