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Quantitative Real-Time PCR: Fluorescent Probe Options and Issues

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Standardization and Quality Assurance in Fluorescence Measurements II

Part of the book series: Springer Series on Fluorescence ((SS FLUOR,volume 6))

Abstract

Fluorescence has played a vital role in the development of polymerase chain reaction (PCR)-based DNA amplification. In qualitative PCR, an end point reaction, the amplified DNA, is visualized using DNA intercalating fluorescent dyes. Creative uses of nucleotide probes with fluorescent tags have been developed for real-time quantitative PCR. These probes take advantage of the behavior and properties of fluorophores. There are advantages and disadvantages to various probe types as well as design considerations. Attention to these issues will help in the development of robust and accurate DNA quantification using real-time PCR.

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Author information

Authors and Affiliations

  1. National Institute of Standards and Technology, Biochemical Science Division, 100 Bureau Drive, Mail Stop 8311, 20899, Gaithersburg, MD, USA

    Marcia J. Holden

  2. National Institute of Standards and Technology, Biochemical Science Division, 100 Bureau Drive, Mail Stop 8312, 20899, Gaithersburg, MD, USA

    Lili Wang

Authors
  1. Marcia J. Holden
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  2. Lili Wang
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Corresponding author

Correspondence to Marcia J. Holden .

Editor information

Ute Resch-Genger

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© 2008 Springer-Verlag Berlin Heidelberg

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Holden, M.J., Wang, L. (2008). Quantitative Real-Time PCR: Fluorescent Probe Options and Issues. In: Resch-Genger, U. (eds) Standardization and Quality Assurance in Fluorescence Measurements II. Springer Series on Fluorescence, vol 6. Springer, Berlin, Heidelberg. https://doi.org/10.1007/4243_2008_046

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