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Quantitation of mRNA by Polymerase Chain Reaction

Nonradioactive PCR Methods

  • Book
  • © 1995

Overview

Authors:
  1. Thomas Köhler

    1. Department of Medicine Institute of Clinical Chemistry and Pathological Biochemistry Molecular Biological Laboratory, University of Leipzig, Leipzig, Deutschland

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  2. Dirk Laßner

    1. Department of Medicine Institute of Clinical Chemistry and Pathological Biochemistry Molecular Biological Laboratory, University of Leipzig, Leipzig, Deutschland

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  3. Harald Remke

    1. Department of Medicine Institute of Clinical Chemistry and Pathological Biochemistry Molecular Biological Laboratory, University of Leipzig, Leipzig, Deutschland

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  4. Anne-Katrin Rost

    1. Department of Medicine Institute of Clinical Chemistry and Pathological Biochemistry Molecular Biological Laboratory, University of Leipzig, Leipzig, Deutschland

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  5. Barbara Thamm

    1. Department of Medicine Institute of Human Genetics Researach Laboratory, University of Leipzig, Leipzig, Deutschland

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  6. Barbara Pustowoit

    1. Department of Medicine Institute of Virology, University of Leipzig, Leipzig, Deutschland

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

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Table of contents (14 chapters)

  1. Front Matter

    Pages I-XX
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  2. Theoretical and Methodical Prerequisities for Using PCR to Quantitate Nucleic Acids

    1. Front Matter

      Pages 1-1
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    2. General Aspects and Chances of Nucleic Acid Quantitation by PCR

      • Th. Köhler
      Pages 3-14

    3. Design of Suitable Primers and Competitor Fragments for Quantitative PCR

      • Th. Köhler
      Pages 15-26

    4. Cloning of Short DNA Fragments and In Vitro Transcription to Generate RNA Standards

      • D. Lassner
      Pages 27-38

    5. Direct Non-Isotopic Sequencing of PCR Products or Standards

      • B. Thamm
      Pages 39-51

  3. Conventional Techniques for mRNA Analysis

    1. Front Matter

      Pages 53-53
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    2. Isolation of mRNA

      • D. Lassner
      Pages 55-63

    3. Synthesis of cDNA

      • D. Lassner
      Pages 65-70

    4. Qualitative RT-PCR: Amplification of Synthesized mdr-1 cDNA

      • Th. Köhler
      Pages 71-80

    5. Single-Tube RT-PCR

      • D. Lassner
      Pages 81-84

    6. Nonradioactive Determination of PCR-Products by Using a DIG Labeled DNA Probe (Dot Blot)

      • Th. Köhler
      Pages 85-91

    7. Nonradioactive Northern Blot Hybridization with DIG-Labeled DNA Probes

      • A.-K. Rost
      Pages 93-114

  4. Semiquantitative and Quantitative Protocols for Measurement of Nucleic Acids by PCR

    1. Front Matter

      Pages 115-115
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    2. Quantitation of mRNA by the ELOSA Technique Using External Standards

      • D. Lassner
      Pages 117-123

    3. Semiquantitative Detection of Viral DNA, e.g. for CMV, by Using the DNA Enzyme Immunoassay (DEIA)

      • B. Pustowoit
      Pages 125-133

    4. HPLC — Analysis of Nucleic Acids

      • H. Remke, Th. Köhler
      Pages 135-141

    5. Quantitation of Absolute Numbers of mRNA Copies in a cDNA Sample by Competitive PCR

      • Th. Köhler
      Pages 143-153

  5. Back Matter

    Pages 155-165
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Keywords

About this book

In this laboratory "cook-book", the authors provide a concise guide to PCR-based techniques to quantify nucleic acids in biological and clinical samples using exclusively nonradioactive detection methods, e.g. HPLC, biotin and digoxigenin based protocols. Each method presentation also includes sections on theory, reagents, standards, applicability, limitations, and trouble shooting. In addition to the protocols, the authors also provide the necessary information on: general aspects of nucleic acid quantitation; design of PCR standards; mRNA purification; cDNA synthesis; solution hybridization; DNA sequencing. This laboratory guide enables professionals as well as beginners to adopt easily quantitative PCR protocols into their own clinical or biomedical research.

Reviews

"...this technical book will be very useful for all customers of PCR. And they are numerous!... Summarizing: an excellent booklet, which we recommend highly and absolutely necessary to all practicians of PCR products and their applications." Cellular and Molecular Biology

Authors and Affiliations

  • Department of Medicine Institute of Clinical Chemistry and Pathological Biochemistry Molecular Biological Laboratory, University of Leipzig, Leipzig, Deutschland

    Thomas Köhler, Dirk Laßner, Harald Remke, Anne-Katrin Rost

  • Department of Medicine Institute of Human Genetics Researach Laboratory, University of Leipzig, Leipzig, Deutschland

    Barbara Thamm

  • Department of Medicine Institute of Virology, University of Leipzig, Leipzig, Deutschland

    Barbara Pustowoit

Bibliographic Information

  • Book Title: Quantitation of mRNA by Polymerase Chain Reaction

  • Book Subtitle: Nonradioactive PCR Methods

  • Authors: Thomas Köhler, Dirk Laßner, Harald Remke, Anne-Katrin Rost, Barbara Thamm, Barbara Pustowoit

  • Series Title: Springer Lab Manuals

  • DOI: https://doi.org/10.1007/978-3-642-79712-5

  • Publisher: Springer Berlin, Heidelberg

  • eBook Packages: Springer Book Archive

  • Copyright Information: Springer-Verlag Berlin Heidelberg 1995

  • Softcover ISBN: 978-3-642-79714-9Published: 29 December 2011

  • eBook ISBN: 978-3-642-79712-5Published: 06 December 2012

  • Edition Number: 1

  • Number of Pages: XX, 166

  • Number of Illustrations: 17 b/w illustrations

  • Topics: Biochemistry, general, Analytical Chemistry, Human Genetics

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