Preface 1 INTRODUCTION TO PCR 1·Setting Up a PCR Laboratory, Theodore E. Mifflin 2·Dealing with Carryover Contamination in PCR: An Enzymatic Strategy, Kyusung Park and Jun Lee 3·High-Fidelity PCR Enzymes, Holly H. Hogrefe and Michael C. Borns 4·Optimization and Troubleshooting in PCR, Kenneth H. Roux 5·PCR Amplification of Highly GC-rich Regions, Lise Lotte Hansen and Just Justesen 6·Tips for Long and Accurate PCR, Wayne M. Barnes 7·PCR Primer Design, Arun Apte and Saurabha Daniel 8·Nonradioactive Cycle Sequencing of PCR-amplified DNA, Susan R. Haynes 2 SAMPLE PREPARATION 9·DNA Purification, Craig Smith, Paul Otto, Rex Bitner, and Gary Shiels 10·RNA Purification, Michael A. Connolly, Peter A. Clausen, and James G. Lazar 11·In Situ Localization of Gene Expression Using Laser Capture Microdissection, Motoko Morimoto, Masahiro Morimoto, Jeannette Whitmire, Robert A. Star, Joseph F. Urban, Jr., and William C. Gause 12·Strategies for Overcoming PCR Inhibition, Peter Radstrom, Charlotta Lofstrom, Maria Lovenklev, Rickard Knutsson, and Petra Wolffs 3 RT-PCR METHODS 13·The Use of Internal Controls for Relative and Competitive Quantitative RT-PCR, Rense M. Horner 14·Comparison of Four Real-time PCR Detection Systems: Bio-Rad I-Cycler, ABI 7700, Roche LightCycler, and the Cepheid Smartcycler, Katherine E. Templeton and Eric C.J. Claas 15·New Technology for Quantitative PCR, Wolfgang Kusser, Sandrine Javorschi, and Martin A. Gleeson 16·Amplification of RNA: High-temperature Reverse Transcription and DNA Amplification with a Magnesium-activated Thermostable DNA Polymerase, Edward S. Smith, Alvin K. Li, Alice M. Wang, David H. Gelfand, and Thomas W. Myers 4 DETECTION OF PCR PRODUCTS: SPECIALIZED APPLICATIONS 17·Loss of Heterozygosity, a Multiplex PCR Method to Define Narrow Deleted Chromosomal Regions of a Tumor Genome, Lise Lotte Hansen and Just Justesen 18·Single-strand Conformation Polymorphism Analysis, Anne E. Jedlicka and Steven R. Kleeberger 19·Reverse Transcriptase In Situ PCR, Gerard J. Nuovo 20·Sensitive and Fast Mutation Detection by Solid-phase Chemical Cleavage of Mismatches, Lise Lotte Hansen, Just Justesen, Andreana Lambrinakos, and Richard G.H. Cotton 5 ANALYSIS OF DIFFERENTIAL GENE EXPRESSION BY PCR 21·Applying PCR for Microarray-based Gene Expression Analysis, Joan M. Zakel 22·Identification of Differential Genes by Suppression Subtractive Hybridization, Denis V. Rebrikov 23·Fluorescent mRNA Differential Display in Gene Expression Analysis, Jeffrey S. Fisher, Jonathan Meade, Jamie Walden, Zhen Guo, and Peng Liang 6 CLONING BY PCR 24·PCR-based Method for Screening Libraries, David I. Israel 25·Beyond Classic RACE （Rapid Amplification of cDNA Ends）, Michael A. Frohman 26·Synthesis and Analysis of DNA Libraries Using PCR, 391 John L. Joyal, Jinwei Jiang, and David I. Israel 7 CLONING OF PCR PRODUCTS 27·Strategies for Cloning PCR Products, Peter D'Arpa 28·Bidirectional and Directional Cloning of PCR Products, Gina L. Costa and Michael P. Weiner 29·Ribocloning: DNA Cloning and Gene Construction Using PCR Primers Terminated with a Ribonucleotide, Wayne M. Barnes 8 MUTAGENESIS BY PCR 30·Mutagenic PCR, R. Craig Cadwell and Gerald F. Joyce 31·Rapid PCR Site-directed Mutagenesis, Michael P. Weiner and Gina L. Costa 32·Mutagenesis and Synthesis of Novel Recombinant Genes Using PCR, Abbe N. Vallejo, Robert J. Pogulis, and Larry R. Pease 33·Streamlined Gene Assembly PCR, Wayne M. Barnes and Elaine R. Frawley APPENDICES 1·Licensing 2·Modification of Oligonucleotides for Use in PCR 3·Cautions 4·Suppliers Index
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