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In the decade since the publication of the bestselling first edition, Polymerase Chain Reaction (PCR) technology has been at the forefront of dramatic developments in biochemistry. Many exciting advances have been made in this specialized field of DNA analysis and a large number of commercial kits and additives are now available. There has never been a better time for a comprehensive resource covering the current range of PCR strategies and uses.
PCR Technology: Current Innovations, Second Edition is a fundamental reference for scientists new to PCR technology and a source of up-to-date applications for those familiar with the method. It provides theoretical considerations, discussions, and a selection of state-of-the-art techniques for mutation studies, clinical diagnosis, and the detection of food-borne pathogens. The book begins with discussions of the preparation of PCR experiments, followed by examples of analytical PCR divided into qualitative and quantitative applications. The final section explores preparative methods addressing DNA generation for further analysis and in vitro evolution. Featuring detailed presentations of PCR protocols, this volume contains critical information for practitioners in a wide variety of fields, including forensics, molecular biology research, clinical DNA diagnostics, botany, and paleontology. It provides the essential tools for developing innovative approaches in this leading technology of the genomics revolution.
SECTION I. GENERAL CONSIDERATIONSTEMPLATE ISOLATION AND PREPARATIONExtraction and Amplification of Ancient Data, Anne C. StoneLong PCR from Damaged DNA, Bernard Fromenty, PhilippeSchmitt, Dominique PessayreQuantitative mRNA Analysis in Small Cell Samples by RT-PCRand Flow Cytometry, Veronika Stemme, Sten StemmeMicrodissection of Single or Small Numbers of Cells for Analysesof DNA and RNA by PCR, Rose G. Mage, Robert F. Bonner, Harold ObiakorPCR OPTIMIZATIONPre-PCR Processing Strategies, Peter R?dstr/m, MariaL/venklev, Petra Wolffs, Charlotta L/fstr/m and RickardKnutssonNovel PCR-Enhancing Compounds and their Modes of Action, RayChakrabartiOptimizing PCR with the Aid of Experimental Design, ThomasWeissensteinerLABELINGEconomic Fluorescent Labeling of PCR-Products forMicrosatellite- and Single-Stranded-Conformation-Polymorphism (SSCP) Analysis, Markus SchuelkeCLONINGUniversal Restriction Site-Free Cloning Method Using Chimeric Primers,Guo Jun ChenSECTION II. ANALYTICAL APPLICATIONSDETECTION OF NUCLEIC ACID VARIATIONHigh-Throughput Genotyping and the PCR, Kalim U. Mir, JiannisRagoussisHigh Throughput Genetic Analysis Through Multiplexed PCR and Multi-Capillary Electrophoresis, John M Butler, Peter M. ValloneThe Good Assay: A Purification Free Assay for Genotyping by Maldi MassSpectrometry, J/rg Tost, Ram n Kucharzak, Doris Lechner, Ivo Glynne GutPrimer Design for Large Scale Multiplex PCR and Arrayed PrimerExtension (APEX), Maido Remm, Ants Kurg, Andres MetspaluSurface Plasmon Resonance Based Biosensor Technology for Real-Time Detection of PCR Products, Giordana Feriotto, Roberto GambariFluorescent Amplified Fragment Length Polymorphism (FAFLP)Genotyping - Catherine ArnoldGlobal Analysis of DNA Allelic Variations (GADAV) by Specific Enrichmentof Mismatches and Selective Amplification of Heterohybrids, XinghuaPan, Sherman M. WeissmanQuantitative DNA-Methylation Analysis by the Bisulfite ConversionMethod, Osman El-Maarri, Manfred Kuepper, Johannes Oldenburg, J/rnWalterSEQUENCINGPyrosequencing Technology, P?l Nyr?n, Tommy Nordstr/m, BabackGharizadehOctamer-Primed Cycle Sequencing, Anelia I. Kraltcheva, Susan H. HardinIN SITU ANALYSISIn Situ Amplification of cDNA, Gerard J. NuovoDetection of a Single Copy Sequence on Human by Cyclic Prins, KirstenMennicke, Sven Opitz, Eberhard SchwingerQUANTITATIVE ANALYSISMeaningful Quantification of mRNA Using Real-Time PCR, Stephen ABustinAbsolute Quantification of Specific Nucleic Acids by (RT)-PCR Using aNonlinear Mathematical Model for Data Analysis, Huong L.Vu, SergeTroubetzkoy, Vu Q. Nguyen, Michael W. Russell, Jiri MesteckySingle-Molecule PCR - Basic Protocols and Applications, Paul H. DearGene Expression and Discovery: Application of a Genome-WideApproach, Timothy A. Lohret, Karen M. KellyThe Detection of Differential Gene Expression Changes Using SAG