PCR Cloning Protocols

From Molecular Cloning to Genetic Engineering

  • Bruce A. White

Part of the Methods in Molecular Biology™ book series (MIMB, volume 67)

Table of contents

  1. Front Matter
    Pages i-xiv
  2. Performing and Optimizing PCR

    1. Front Matter
      Pages 1-1
    2. PCR
      Lori A. Kolmodin, J. Fenton Williams
      Pages 3-15
    3. Suzanne Cheng, Lori A. Kolmodin
      Pages 17-29
    4. Kenneth H. Roux, Karl H. Hecker
      Pages 39-45
    5. Alain Moreau, Colette Duez, Jean Dusart
      Pages 47-53
  3. Cloning PCR Products

    1. Front Matter
      Pages 61-61
    2. Alan R. Shuldiner, Keith Tanner
      Pages 69-78
    3. Robert M. Horton, Raghavanpillai Raju, Bianca M. Conti-Fine
      Pages 101-110
    4. Gustavo Caetano-Anollés, Robert N. Trigiano
      Pages 111-127
  4. Mutagenesis, Recombination, and In Vitro Selection

    1. Front Matter
      Pages 129-129
    2. Douglas H. Jones, Stanley C. Winistorfer
      Pages 131-140
    3. Robert M. Horton
      Pages 141-150

About this book

Introduction

The advent of PCR, with its power to amplify tiny amounts of DNA, quickly spawned the development of many analytical procedures that are widely used for detection, measurement, and characterization. However, creative investigators soon discovered the power of PCR for synthetic or preparative uses. This volume focuses on such preparative PCR protocols, which can be used in the cloning and modification of DNA. PCR Cloning Protocols: From Molecular Cloning to Genetic Engineer­ ing is divided into seven parts, each containing a collection of chapters address­ ing a general approach or goal. Part I presents basic PCR protocols, emphasizing optimizing conditions for (he amplification of DNA fragments of several kilobases in length. Part 11 offers several procedures for cloning PCR prod­ ucts, depending on whether a specific restriction site can be used in the clon­ ing vector, the PCR product is to be gel purified before cloning, or the fragmeni needs to be inserted in one or both orientations. Part III includes several pro­ tocols involved in the mutagenesis of DNA, either site-directed or not, as well as several approaches to recombinant PCR, either for mutagenesis or building a custom gene, as well as one chapter describing a specific use of in vitro selection. Part IV addresses the frequent need to amplify and clone segments of DNA that are to the right, left, or scattered within a stretch of DNA (either vector, chromosomal, or cDNA) of known sequence.

Editors and affiliations

  • Bruce A. White
    • 1
  1. 1.Department of AnatomyUniversity of Connecticut Health CenterFarmington

Bibliographic information

  • DOI https://doi.org/10.1385/0896034836
  • Copyright Information Humana Press 1997
  • Publisher Name Humana Press, Totowa, NJ
  • eBook Packages Springer Protocols
  • Print ISBN 978-0-89603-483-9
  • Online ISBN 978-1-59259-553-2
  • Series Print ISSN 1064-3745
  • Series Online ISSN 1940-6029
  • About this book