PCR Cloning Protocols

ISBN-13: 9781617372810 / Angielski / Miękka / 2010 / 439 str.

Bing-Yuan Chen; Harry W. Janes

Opis

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.

Recenzje

"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

Spis treści

Part I. Performing and Optimizing PCR Polymerase Chain Reaction: Basic Principles and Routine Practice Lori A. Kolmodin and David E. Birch Computer Programs for PCR Primer Design and Analysis Bing-Yuan Chen, Harry W. Janes, and Steve Chen Single-Step PCR Optimization Using Touchdown and Stepdown PCR Programming Kenneth H. Roux XL PCR Amplification of Long Targets from Genomic DNA Lori A. Kolmodin Coupled One-Step Reverse Transcription and Polymerase Chain Reaction Procedure for Cloning Large cDNA Fragments Jyrki T. Aatsinki Long Distance Reverse-Transcription PCR Volker Thiel, Jens Herold, and Stuart G. Siddell Increasing PCR Sensitivity for Amplification from Paraffin-Embedded Tissues Abebe Akalu and Juergen K. V. Reichardt GC-Rich Template Amplification by Inverse PCR: DNA Polymerase and Solvent Effects Alain Moreau, Da Shen Wang, Steve Forget, Colette Duez, and Jean Dusart PCR Procedure for the Isolation of Trinucleotide Repeats Teruaki Tozaki Methylation-Specific PCR Haruhiko Ohashi 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, and Xuetao Cao Part II. Cloning PCR Products Cloning PCR Products: An Overview Baotai Guo and Yuping Bi Using T4 DNA Polymerase to Generate Clonable PCR Products Kai Wang Enzyme-Free Cloning of PCR Products and Fusion Protein Expression Brett A. Neilan and Daniel Tillett Directional Restriction Site-Free Insertion of PCR Products into Vectors Guo Jun Chen Autosticky PCR: Directional Cloning of PCR Products with Preformed 5' Overhangs József Gál and Miklós Kálmán A Rapid and Simple Procedure for Direct Cloning of PCRProducts into Baculoviruses Tamara S. Gritsun, Michael V. Mikhailov, and Ernest A. Gould Part III. Mutagenesis and Recombination PCR Approaches to DNA Mutagenesis and Recombination: An Overview Binzhang Shen In-Frame Cloning of Synthetic Genes Using PCR Inserts James C. Pierce Megaprimer PCR Sailen Barik PCR-Mediated Recombination: A General Method Applied to Construct Chimeric Infectious Molecular Clones Guowei Fang, Barbara Weiser, Aloise Visosky, Timothy Moran, and Harold Burger PCR Method for Generating Multiple Mutations at Adjacent Sites Jiri Adamec A Fast Polymerase Chain Reaction-Mediated Strategy for Introducing Repeat Expansions into CAG-Repeat Containing Genes Franco Laccone PCR Screening in Signature-Tagged Mutagenesis of Essential Genes Dario E. Lehoux and Roger C. Levesque Staggered Extension Process (StEP) In Vitro Recombination Anna Marie Aguinaldo and Frances Arnold Random Mutagenesis by Whole-Plasmid PCR Amplification Donghak Kim and F. Peter Guengerich Part IV. Cloning Unknown Neighboring DNA PCR-Based Strategies to Clone Unknown DNA Regions from Known Foreign Integrants: An Overview Eric Ka-Wai Hui, Po-Ching Wang, and Szecheng J. Lo Long Distance Vectorette PCR (LDV PCR) James A. L. Fenton, Guy Pratt, and Gareth J. Morgan Nonspecific, Nested Suppression PCR Method for Isolation of Unknown Flanking DNA ('Cold-Start Method') Michael Lardelli Inverse PCR: cDNA Cloning Sheng-He Huang Inverse PCR: Genomic DNA Cloning Ambrose Y. Jong, Anna T'ang, De-Pei Liu, and Sheng-He Huang Gene Cloning and Expression Profiling by Rapid Amplification of Gene Inserts with Universal Vector Primers Sheng-He Huang, Hua-Yang Wu, and Ambrose Y. Jong The Isolation of DNA Sequences Flanking Tn5 Trans

Blurb

In the post-genomic era, PCR has become the method of choice not only for cloning existing genes, but also for generating a wide array of novel genes by mutagenesis and/or recombination within the genes of interest. 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. Powerful applications of PCR in library construction and sublibrary generation and screening are also presented.
Authoritative and up-to-date, PCR Cloning Protocols, Second Edition, constitutes a gold-standard collection of the fastest, simplest, and most popular methods for isolating genes from all biological samples and creating novel genes from them by mutagenesis/recombination-essential methods for today's study of functional genomics, gene expression, protein structure-function relationships, protein engineering, and molecular evolution.