PCR Protocols

Drawing on the highly successful first edition, this newly-revised second edition covers the many advances made in PCR technology since the first book, which has been used in more than 10,000 laboratories worldwide. As PCR technology has advanced significantly since the first edition, and has expanded its use in the clinical laboratory of physician/researchers, the scope of this book is greatly expanded to enable researchers at all levels to easily reproduce and adapt PCR experiments to their own specific requirements. The meethods selected represent worked examples from many fields that can be reproduced and adapted for use within the reader's laboratory. The authors have provided both a primer to allow the reader to gain basic experience of different PCR techniques, as well as in-depth insight into a variety of the more complex applications of PCR. This book will be essential for the labs of all biochemists, molecular biologists, geneticists and researchers utilizing the PCR techinque in their work.

Part I. Introduction to PCR A Short History of the Polymerase Chain Reaction John M. S. Bartlett and David Stirling PCR Patent Issues Peter Carroll and David Casimir Equipping and Establishing a PCR Laboratory Susan McDonagh Quality Control in PCR David Stirling Part II. Preparation of Nucleic Acid Templates Extraction of Nucleic Acid Templates John M. S. Bartlett Extraction of DNA from Whole Blood John M. S. Bartlett and Anne White DNA Extraction from Tissue Helen Pearson and David Stirling Extraction of DNA from Microdissected Archival Tissues James J. Going RNA Extraction from Blood Helen Pearson RNA Extraction from Frozen Tissue John M. S. Bartlett RNA Extraction from Tissue Sections Helen Pearson Dual DNA/RNA Extraction David Stirling and John M. S. Bartlett DNA Extraction from Fungi, Yeast, and Bacteria David Stirling Isolation of RNA Viruses from Biological Materials Susan McDonagh Extraction of Ancient DNA Wera M. Schmerer DNA Extraction from Plasma and Serum David Stirling Technical Notes for the Detection of Nucleic Acids John M. S. Bartlett Technical Notes for the Recovery and Purification of PCR Products from Acrylamide Gels David Stirling Part III. Basic PCR Methods PCR Primer Design David L. Hyndman and Masato Mitsuhashi Optimization of Polymerase Chain Reactions Haiying Grunenwald Subcycling PCR for Long-Distance Amplifications of Regions with High and Low Guanine-Cystine Content: Amplification of the Intron 22 Inversion of the FVIII Gene David Stirling Rapid Amplification of cDNA Ends Xin Wang and W. Scott Young III Randomly Amplified Polymorphic DNA Fingerprinting: The Basics Ranil S. Dassanayake and Lakshman P. Samaranayake Microsphere-Based Single Nucleotide Polymorphism Genotyping Marie A. Iannone, J. David Taylor, Jingwen Chen, May-Sung Li, Fei Ye, and Michael P. Weiner Ligase Chain Reaction William H. Benjamin, Jr., Kim R. Smith, and Ken B. Waites Nested RT-PCR in a Single Closed Tube Antonio Olmos, Olga Esteban, Edson Bertolini, and Mariano Cambra Direct PCR from Serum: Application to Viral Genome Detection Kenji Abe Long PCR Amplification of Large Fragments of Viral Genomes: A Technical Overview Raymond Tellier, Jens Bukh, Suzanne U. Emerson, and Robert H. Purcell Long PCR Methodology Raymond Tellier, Jens Bukh, Suzanne U. Emerson, and Robert H. Purcell Part IV. Ultrasensitive and Quantitative PCR Qualitative and Quantitative PCR: A Technical Overview David Stirling Ultrasensitive PCR Detection of Tumor Cells in Myeloma Friedrich W. Cremer and Marion Moos Ultrasensitive Quantitative PCR to Detect RNA Viruses Susan McDonagh Quantitative PCR for cAMP RI Alpha mRNA: Use of Site-Directed Mutation and PCR Mimics John M. S. Bartlett Quantitation of Multiple RNA Species Ron Kerr Part V. Transcriptome Analysis Differential Display: A Technical Overview John M. S. Bartlett AU-Differential Display, Reproducibility of a Differential mRNA Display Targeted to AU Motifs Orlando Dominguez, Lidia Sabater, Yaqoub Ashhab, Eva Belloso, and Ricardo Pujol-Borrell PCR Fluorescence Differential Display Kostya Khalturin, Sergej Kuznetsov, and Thomas C. G. Bosch Microarray Analysis Using RNA Arbitrarily Primed PCR Steven Ringquist, Gaelle Rondeau, Rosa-Ana Risques, Takuya Higashiyama, Yi-Peng Wang, Steffen Porwollik, David Boyle, Michael McClelland, and John Welsh Oligonucleoti