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PCR Cloning Protocols

  • Book
  • © 2002

Overview

Editors:
  1. Bing-Yuan Chen

    1. Department of Plant Science, Rutgers University, New Brunswick

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    You can also search for this editor in PubMed   Google Scholar

  2. Harry W. Janes

    1. Department of Plant Science, Rutgers University, New Brunswick

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    You can also search for this editor in PubMed   Google Scholar

Part of the book series: Methods in Molecular Biology (MIMB, volume 192)

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Table of contents (44 protocols)

  1. Front Matter

    Pages i-xiv
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  2. Performing and Optimizing PCR

    1. Front Matter

      Pages 1-108
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    2. Polymerase Chain Reaction

      • Lori A. Kolmodin, David E. Birch
      Pages 3-18

    3. Computer Programs for PCR Primer Design and Analysis

      • Bing-Yuan Chen, Harry W. Janes, Steve Chen
      Pages 19-29

    4. Single-Step PCR Optimization Using Touchdown and Stepdown PCR Programming

      • Kenneth H. Roux
      Pages 31-36

    5. XL PCR Amplification of Long Targets from Genomic DNA

      • Lori A. Kolmodin
      Pages 37-51

    6. Coupled One-Step Reverse Transcription and Polymerase Chain Reaction Procedure for Cloning Large cDNA Fragments

      • Jyrki T. Aatsinki
      Pages 53-58

    7. Long Distance Reverse-Transcription PCR

      • Volker Thiel, Jens Herold, Stuart G. Siddell
      Pages 59-66

    8. Increasing PCR Sensitivity for Amplification from Paraffin-Embedded Tissues

      • Abebe Akalu, Juergen K. V. Reichardt
      Pages 67-74

    9. GC-Rich Template Amplification by Inverse PCR

      • Alain Moreau, Da Shen Wang, Steve Forget, Colette Duez, Jean Dusart
      Pages 75-80

    10. PCR Procedure for the Isolation of Trinucleotide Repeats

      • Teruaki Tozaki
      Pages 81-89

    11. Methylation-Specific PCR

      • Haruhiko Ohashi
      Pages 91-97

    12. Direct Cloning of Full-Length Cell Differentially Expressed Genes by Multiple Rounds of Subtractive Hybridization Based on Long-Distance PCR and Magnetic Beads

      • Xin Huang, Zhenglong Yuan, Xuetao Cao
      Pages 99-108

  3. Cloning PCR Products

    1. Front Matter

      Pages 109-163
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    2. Cloning PCR Products

      • Baotai Guo, Yuping Bi
      Pages 111-119

    3. Using T4 DNA Polymerase to Generate Clonable PCR Products

      • Kai Wang
      Pages 121-124

    4. Enzyme-Free Cloning of PCR Products and Fusion Protein Expression

      • Brett A. Neilan, Daniel Tillett
      Pages 125-132

    5. Directional Restriction Site-Free Insertion of PCR Products into Vectors

      • Guo Jun Chen
      Pages 133-139

    6. Autosticky PCR

      • József Gál, Miklós Kálmán
      Pages 141-151

    7. A Rapid and Simple Procedure for Direct Cloning of PCR Products into Baculoviruses

      • Tamara S. Gritsun, Michael V. Mikhailov, Ernest A. Gould
      Pages 153-163
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Keywords

About this book

PCR Cloning Protocols, Second Edition, updates and expands Bruce White's best-selling PCR Cloning Protocols (1997) with the newest procedures for DNA cloning and mutagenesis. Here the researcher will find readily reproducible methods for all the major aspects of PCR use, including PCR optimization, computer programs for PCR primer design and analysis, and novel variations for cloning genes of special characteristics or origin, with emphasis on long distance PCR and GC-rich template amplification. Also included are both conventional and novel enzyme-free and restriction site-free procedures to clone PCR products into a range of vectors, as well as state-of-the-art protocols to facilitate DNA mutagenesis and recombination, and to clone the challenging uncharacterized DNA flanking a known DNA fragment.

Reviews

"This hardcover book is packed full of detailed protocols and methodological articles on various aspects of PCR. The fact that the book was taken out of my hands by a colleague, to read up on touch-down PCR, as soon as I received it for review, attest to the book's usefulness. I will have to chain this book to my bookshelf, to make sure it does not disappear quickly-or I may consider buying a couple for the laboratory." -Plant Science

Review of the first edition.
This volume provides protocols useful to both the inexperienced and the experienced investigator. Most importantly it highlights the emerging role of PCR as a versatile tool for the molecular biologist eliminating many laborious and expensive techniques associated with traditional gene isolation and analysis.-Biochemical Education

Editors and Affiliations

  • Department of Plant Science, Rutgers University, New Brunswick

    Bing-Yuan Chen, Harry W. Janes

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