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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