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Protocols for Nucleic Acid Analysis by Nonradioactive Probes

  • Book
  • © 2007

Overview

Editors:
  1. Elena Hilario

    1. Gene Technologies, HortResearch Ltd., Mt. Albert, Auckland, New Zealand

    View editor publications

    You can also search for this editor in PubMed   Google Scholar

  2. John Mackay

    1. Applied Science, Roche Diagnostics, Auckland, New Zealand

    View editor publications

    You can also search for this editor in PubMed   Google Scholar

  • Readily reproducible protocols for the analysis of nucleic acids by nonradioactive methods
  • New fluorescent techniques such as Real Time PCR and microarrays
  • Follow step-by-step instructions that ensure successful results
  • Master tricks of the trade, troubleshoot, and avoid known pitfalls
  • Includes supplementary material: sn.pub/extras

Part of the book series: Methods in Molecular Biology (MIMB, volume 353)

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Table of contents (19 protocols)

  1. Front Matter

    Pages i-x
    Download chapter PDF

  2. Nucleic Acid Extractions

    1. Front Matter

      Pages 1-1
      Download chapter PDF

    2. Genomic DNA Isolation From Different Biological Materials

      • Duckchul Park
      Pages 3-13

    3. Extraction of Plant RNA

      • Elspeth MacRae
      Pages 15-24

  3. Indirect Detection

    1. Front Matter

      Pages 25-25
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    2. Overview of Hybridization and Detection Techniques

      • Elena Hilario
      Pages 27-38

    3. Checkerboard DNA-DNA Hybridization Technology Using Digoxigenin Detection

      • Lisa S. Gellen, Glenn M. Wall-Manning, Chris H. Sissons
      Pages 39-67

    4. Nonradioactive Northern and Southern Analyses From Plant Samples

      • Christoph Peterhaensel, Dagmar Weier, Thomas Lahaye
      Pages 69-78

    5. Screening a BAC Library With Nonradioactive Overlapping Oligonucleotide (Overgo) Probes

      • Elena Hilario, Tiffany F. Bennell, Erik Rikkerink
      Pages 79-91

    6. Direct In-Gel Hybridization of DNA With Digoxigenin-Labeled Probes

      • Saeed A. Khan, Mohamed S. Nawaz
      Pages 93-103

    7. In Situ Hybridization of Termite Microbes

      • Shigeharu Moriya, Satoko Noda, Moriya Ohkuma, Toshiaki Kudo
      Pages 105-112

  4. Fluorescent Labeling and Detection

    1. Front Matter

      Pages 113-113
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    2. RNA Electrophoretic Mobility Shift Assay Using a Fluorescent DNA Sequencer

      • Yukinori Eguchi
      Pages 115-124

    3. Comparative Quantitation of mRNA Expression in the Central Nervous System Using Fluorescence In Situ Hybridization

      • Darren J. Day, Eli M. Mrkusich, John H. Miller
      Pages 125-142

    4. Visualization of Gene Expression by Fluorescent Multiplex Reverse Transcriptase-PCR Amplification

      • María Rosa Ponce, Víctor Quesada, Andrea Hricová, José Luis Micol
      Pages 143-152

    5. Fluorescence In Situ Hybridization for the Identification of Environmental Microbes

      • Annelie Pernthaler, Jakob Pernthaler
      Pages 153-164

  5. Kinetic (“Real-Time”) PCR

    1. Front Matter

      Pages 165-165
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    2. Introduction to Kinetic (Real-Time) PCR

      • John Mackay
      Pages 167-176

    3. Validation of Short Interfering RNA Knockdowns by Quantitative Real-Time PCR

      • Sukru Tuzmen, Jeff Kiefer, Spyro Mousses
      Pages 177-203

    4. Real-Time Quantitative PCR as an Alternative to Southern Blot or Fluorescence In Situ Hybridization for Detection of Gene Copy Number Changes

      • Jasmien Hoebeeck, Frank Speleman, Jo Vandesompele
      Pages 205-226
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Keywords

About this book

Protocols for Nucleic Acid Analysis by Non-radioactive Probes, Second Edition provides a firm background on the basic preparative protocols required for the analysis of nucleic acids by nonradioactive methods. Presenting the methodologies using amazing new applications, this volume offers guide chapters on nucleic acid extractions, preparation of nucleic acid blots, and labeling of nucleic acids with nonradioactive haptens. New fluorescent techniques such as Real Time PCR and microarrays are also included, allowing users to get a nonradioactive protocol implemented in the laboratory with minimum adaptation required and fastest time to results.
The protocols follow the successful Methods in Molecular Biology™ series format, each offering step-by-step laboratory instructions, an introduction outlining the principles behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.

Reviews

From the reviews of the second edition:

...this book provides a collection of good protocols which are in general clearly written. They can be easily employed on the bench without being a specialist in a particular fields. - FEBS Letters

...Covers the basics of a variety of probe-labelling systems very well...a very interesting read. It could save you money and trouble from the radiation safety officer.-Biotechnology and Applied Biochemistry

"This is a set of protocols for various types of nucleic acid analysis using nonradiocative methods. … The goal is to provide protocols for nucleic acid analysis using other than radioactive reporter systems. This approach is commonplace at present, so a volume of practical utility has merit. … The target audience would be research laboratory personnel working in this general area. The editors have recruited a group of contributors with worldwide representation." (Eugene A Davidson, Doody’s Review Service, November, 2007)

"The editors of this volume have gathered together a variety of protocols for anyone who wishes to use nonradioactive methods for the analysis of their chosen nucleic acid molecules. ... Combined with the excellent overviews on microarrays as well as hybridization and detection techniques, and several useful assays for the application of these techniques for nucleic acid analysis, scientists will find this book handy for their research and educators will adopt it as a useful teaching tool for advanced students." (Tzvi Tzfira, The Quarterly Review of Biology, Vol. 83, 2008)

Editors and Affiliations

  • Gene Technologies, HortResearch Ltd., Mt. Albert, Auckland, New Zealand

    Elena Hilario

  • Applied Science, Roche Diagnostics, Auckland, New Zealand

    John Mackay

Bibliographic Information

  • Book Title: Protocols for Nucleic Acid Analysis by Nonradioactive Probes

  • Editors: Elena Hilario, John Mackay

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1385/1597452297

  • Publisher: Humana Totowa, NJ

  • eBook Packages: Springer Protocols

  • Copyright Information: Humana Press 2007

  • Hardcover ISBN: 978-1-58829-430-2Published: 12 January 2007

  • Softcover ISBN: 978-1-61737-564-4Published: 01 December 2010

  • eBook ISBN: 978-1-59745-229-8Published: 02 February 2008

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 2

  • Number of Pages: 321

  • Number of Illustrations: 61 b/w illustrations, 5 illustrations in colour

  • Topics: Biochemistry, general, Human Genetics, Cell Biology, Plant Sciences

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