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Quantitative PCR Protocols

  • Book
  • © 1999

Overview

Editors:
  1. Bernd Kochanowski

    1. University of Regensburg, Germany

    View editor publications

    You can also search for this editor in PubMed   Google Scholar

  2. Udo Reischl

    1. University of Regensburg, Germany

    View editor publications

    You can also search for this editor in PubMed   Google Scholar

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

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Table of contents (20 protocols)

  1. Front Matter

    Pages i-x
    Download chapter PDF

  2. Reviews

    1. Front Matter

      Pages 1-1
      Download chapter PDF

    2. Quantitative PCR

      • Udo Reischl, Bernd Kochanowski
      Pages 3-30

    3. General Principles of Quantitative PCR

      • Luc Raeymaekers
      Pages 31-41

    4. Effects of Collection, Processing, and Storage on RNA Detection and Quantification

      • Mark Holodniy
      Pages 43-59

    5. Quantitative RT-PCR

      • Paul D. Siebert
      Pages 61-85

    6. Kinetic Quantitative PCR vs End-Point Quantitative PCR with Internal Standard

      • Olivier Lantz, Elizabeth Bonney, Franck Griscelli, Yassine Taoufik
      Pages 87-102

    7. Comparison of Competitive PCR and Positive Control-Based PCR

      • François Mallet
      Pages 103-116

  3. Protocols

    1. Front Matter

      Pages 117-117
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    2. End-Point Titration-PCR for Quantitation of Cytomegalovirus DNA

      • Jerzy K. Kulski
      Pages 119-130

    3. Analysis of Amplified DNA Molecules by Capillary Electrophoresis and Laser Induced Fluorescence

      • Michael J. Fasco
      Pages 131-146

    4. Competitor Calibration and Analysis of Competitive Amplified PCR Products by High-Performance Liquid Chromatography (HPLC)

      • Thomas Köhler
      Pages 147-155

    5. Quantifying Amplicons with ELISA

      • Olivier Lantz, Elizabeth Bonney, Yassine Taoufik
      Pages 157-170

    6. Competitive PCR Quantitation Utilizing a Microtiter Plate Based Format for the Detection of PCR Products

      • Bernd Kochanowski, Wolfgang Jilg
      Pages 171-182

    7. Competitive and Differential RT-PCR (CD-RT-PCR) for Measurement of Normalized Gene Expression Using Antisense Competitors

      • Eric de Kant
      Pages 183-195

    8. Amplified Assay for Specific Dual-Labeled DNA Using the Coagulation Cascade (EDNA-ELCA)

      • George J. Doellgast, G. Alan Beard, Margaret Sheehan, Nathan Iyer, Louis S. Kucera, Stephen H. Richardson
      Pages 197-216

    9. Quantitative PCR with Internal Standardization and OLA-ELISA Product Analysis for the p53 Tumor Suppressor Gene

      • Meinhard Hahn, Alfred Pingoud
      Pages 217-231

    10. Quantitative Analysis of Human DNA Sequences by PCR and Solid-Phase Minisequencing

      • Anu Suomalainen, Ann-Christine Syvänen
      Pages 233-244

    11. High Resolution PCR Quantitation by AmpliSensor Assay

      • Chang-Ning Wang
      Pages 245-251

    12. Construction of Polycompetitors for Competitive PCR

      • David B. Corry, Richard M. Locksley
      Pages 253-264
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About this book

Since the polymerase chain reaction (PCR) was first developed in 1985, an enormous number of research reports have documented the versatility of this brilliant technique for in vitro amplification of nucleic acids. Although PCR has had a profound impact in many areas of research, contrary to exp- tation its routine application to the quantitation of nucleic acids has proven problematic in several aspects. The shortcomings are principally caused by the exponential nature of PCR, whereby small variations in amplification ef- ciency may dramatically affect the yield of amplification product. Even mi- mal temperature deviations that occur between adjacent wells of a thermocycler or day-to-day variations in the efficiency of nucleic acid preparation can lead to significant differences in the extent of amplification between otherwise id- tical samples. However, knowing more about the intrinsic limitations of PCR is the first step towards surmounting the shortcomings associated with this prom- ing methodology. With the introduction of appropriate standards of known amount, which are co-amplified with the sample using the same primers, it is increasingly feasible to address biological or diagnostic questions that are d- ficult or impossible to answer using any other experimental approach.

Reviews

"...The book is very comprehensive and well written..."-The Biochemist

Editors and Affiliations

  • University of Regensburg, Germany

    Bernd Kochanowski, Udo Reischl

Bibliographic Information

  • Book Title: Quantitative PCR Protocols

  • Editors: Bernd Kochanowski, Udo Reischl

  • Series Title: Methods in Molecular Medicine

  • DOI: https://doi.org/10.1385/0896035182

  • Publisher: Humana Totowa, NJ

  • eBook Packages: Springer Protocols

  • Copyright Information: Humana Press 1999

  • eBook ISBN: 978-1-59259-262-3Published: 01 February 2008

  • Series ISSN: 1543-1894

  • Series E-ISSN: 1940-6037

  • Edition Number: 1

  • Number of Pages: 320

  • Topics: Cell Biology

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