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Quantitative Measurements

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Capillary Electrophoresis of Nucleic Acids

Part of the book series: Methods in Molecular Biology ((MIMB,volume 162))

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Abstract

The fundamental role of any separation process is the isolation and quantitation of the individual components of a mixture. In the analysis of DNA by capillary electro-phoresis (CE), the migration time and the sample quantity are the basic parameters obtained from any electropherogram. Migration time is used to estimate the DNA fragment size, and the quantity of product gives the analyst polymerase chain reaction (PCR) reaction yields, the efficiency of an extraction process, or the level of expression of a particular gene. Chemical processes such as PCR amplification or restriction enzyme digestion can vary in their yield, making measurement of the product quantity a critical parameter. The need for exact quantitative measurements is increasing with the use of methods such as quantitative PCR, in which an analyte and an internal standard of similar sequence are amplified together and compared. Many different applications, such as mutational analysis, and the detection of length polymorphisms, require the ability to reliably detect the size of a DNA fragment. DNA typing can require the routine determinations of differences in fragment size of two bases or less.

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Author information

Authors and Affiliations

  1. Department of Chemistry, Clippinger Laboratories, Ohio University, Athens, OH

    Bruce McCord

Authors
  1. Bruce McCord
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Editor information

Editors and Affiliations

  1. Australian Genome Research Facility, University of Queensland, Brisbane

    Keith R. Mitchelson

  2. Walter & Eliza Hall Institute, Parkville, Australia

    Keith R. Mitchelson

  3. Biochip Research and Development Center, State Key Laboratory for Biomembrane and Membrane Biotechnology, Tsinghua University, Beijing, China

    Jing Cheng

  4. Aviva Biosciences Corporation, San Diego, CA

    Jing Cheng

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© 2001 Humana Press Inc.

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McCord, B. (2001). Quantitative Measurements. In: Mitchelson, K.R., Cheng, J. (eds) Capillary Electrophoresis of Nucleic Acids. Methods in Molecular Biology, vol 162. Humana Press. https://doi.org/10.1385/1-59259-055-1:51

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  • DOI: https://doi.org/10.1385/1-59259-055-1:51

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-779-3

  • Online ISBN: 978-1-59259-055-1

  • eBook Packages: Springer Protocols

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