InhaltsangabePart 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 BingYuan Chen, Harry W. Janes, and Steve Chen SingleStep PCR Optimization Using Touchdown and Stepdown PCR Programming Kenneth H. Roux XL PCR Amplification of Long Targets from Genomic DNA Lori A. Kolmodin Coupled OneStep 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 GCRich 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 MethylationSpecific 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 EnzymeFree 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 InFrame Cloning of Synthetic Genes Using PCR Inserts James C. Pierce Megaprimer PCR Sailen Barik PCRMediated 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 PCRBased Strategies to Clone Unknown DNA Regions from Known Foreign Integrants: An Overview Eric KaWai Hui, PoChing 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 ShengHe 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 ShengHe Huang, HuaYang Wu, and Ambrose Y. Jong The Isolation of DNA Sequences Flanking Tn5 Trans