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RT-PCR, the most widely used technique in biomedical research for analyzing gene expression, has proven a significant boon to the molecular investigation of disease pathogenesis. In RT-PCR Protocols, Joe O'Connell has assembled a panel of highly regarded molecular biologists and clinical researchers to describe in detail their most novel, useful, and interesting RT-PCR applications. Here the newcomer will find readily reproducible protocols for highly sensitive detection and quantification of gene expression, the in situ localization of gene expression in tissue, and the cloning of genes, as well as for analyzing T-cell clones and the differential expression of genes. For the expert seeking to extend the usefulness of RT-PCR, there are user-friendly applications that complement the latest technological advances, including laser-capture microdissection (LCM), real-time and quantitative PCR, microarray technology, cDNA cloning, and antibody engineering. Each protocol emphasizes the technical steps critical for experimental success and includes tips on avoiding potential pitfalls, notes explaining the method's scientific basis and its possible sources of error, and comments on how it might best be accommodated to various experimental circumstances. Diverse and wide-ranging, RT-PCR Protocols offers investigators powerful techniques to be used in the molecular investigation of disease pathogenesis and shows how RT-PCR can complement other technological advances in the design of today's new therapeutic strategies.

Polymerase chain reaction --- Réaction en chaîne de la polymérase --- Laboratory manuals. --- Manuels de laboratoire --- Electronic books. -- local. --- Polymerase chain reaction -- Laboratory manuals. --- Polymerization. --- Reverse Transcriptase Polymerase Chain Reaction --- Polymerase Chain Reaction --- Nucleic Acid Amplification Techniques --- Genetic Techniques --- Investigative Techniques --- Analytical, Diagnostic and Therapeutic Techniques and Equipment --- Animal Biochemistry --- Human Anatomy & Physiology --- Health & Biological Sciences --- Réaction en chaîne de la polymérase --- Polymerisation --- Polymers --- Polymers and polymerization --- Synthesis of polymers --- Chemical reactions --- Synthesis --- Cytology. --- Cell Biology. --- Cell biology --- Cellular biology --- Biology --- Cells --- Cytologists --- Polymerase chain reaction - Laboratory manuals

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In the post-genomic era, PCR has become the method of choice not only for cloning existing genes, but also for generating a wide array of novel genes by mutagenesis and/or recombination within the genes of interest. PCR Cloning Protocols, Second Edition, updates and expands Bruce White's best-selling PCR Cloning Protocols (1997) with the newest procedures for DNA cloning and mutagenesis. Here the researcher will find readily reproducible methods for all the major aspects of PCR use, including PCR optimization, computer programs for PCR primer design and analysis, and novel variations for cloning genes of special characteristics or origin, with emphasis on long-distance PCR and GC-rich template amplification. Also included are both conventional and novel enzyme-free and restriction site-free procedures to clone PCR products into a range of vectors, as well as state-of-the-art protocols to facilitate DNA mutagenesis and recombination and to clone the challenging uncharacterized DNA flanking a known DNA fragment. Powerful applications of PCR in library construction and sublibrary generation and screening are also presented. Authoritative and up-to-date, PCR Cloning Protocols, Second Edition, constitutes a gold-standard collection of the fastest, simplest, and most popular methods for isolating genes from all biological samples and creating novel genes from them by mutagenesis/recombination-essential methods for today's study of functional genomics, gene expression, protein structure-function relationships, protein engineering, and molecular evolution.

Molecular cloning --- Polymerase chain reaction --- Clonage moléculaire --- Réaction en chaîne de la polymérase --- Laboratory manuals --- Manuels de laboratoire --- Electronic books. -- local. --- Molecular cloning -- Laboratory manuals. --- Polymerase chain reaction -- Laboratory manuals. --- Genetics --- Biology --- Health & Biological Sciences --- Clonage moléculaire --- Réaction en chaîne de la polymérase --- Life sciences. --- Science. --- Biochemistry. --- Cell biology. --- Life Sciences. --- Biochemistry, general. --- Cell Biology. --- Science, general. --- Cell biology --- Cellular biology --- Cells --- Cytologists --- Biological chemistry --- Chemical composition of organisms --- Organisms --- Physiological chemistry --- Chemistry --- Medical sciences --- Natural science --- Science of science --- Sciences --- Biosciences --- Sciences, Life --- Science --- Composition --- Cloning, Molecular --- DNA cloning --- Gene cloning --- Cloning --- Genetic engineering --- Molecular genetics --- Clone cells --- Science, Humanities and Social Sciences, multidisciplinary. --- Cytology.

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PCR (polymerase chain reaction) is now one of the most widely used of basic molecular biology techniques and is an indispensable research tool for the molecular biologist. The basic PCR technique provides the cornerstone for in vitro DNA amplification - allowing the researcher to produce large quantities of DNA from minute amounts of starting material.PCR: A Practical Approach Volume 2 is not a revised version of PCR: A Pracical Approach, but sets out to address some of the exciting new applications fo PCR including cDNA cloning, mRNA quantitation and expression of proteins from PCR products, genomic DNA mapping and fingerprinting, and mutational analysis. The Polymerase Chain Reaction (PCR) has become one of the most widely used techniques in biomedical research, enabling the fast, inexpensive production of large quantities of DNA from minute amounts of starting material. Although the basic procedure is very simple, many variations of it have been developed for different applications, for use in such diverse areas as forensic and diagnostic medicine, genetic engineering, and the food industry. PCR: A Practical Approach provides a general introduction to PCR for researchers new to the area, followed by chapters detailing specific applications of the technique. It covers gene analysis at the level of restriction enzyme polymorphism, point mutation and sequence analysis. Methods for using different source materials - cloned DNA, genomic DNA, RNA, nucleic acid from archive material and PCR products themselves - are given. The emphasis throughout is on the practical applications of the technique and detailed experimental protocols are included at all stages, backed up by advice on avoiding potential pitfalls. This volume provides a starting point for those new to PCR and will also serve as a useful reference for more experienced researchers in all areas of biological science.

General biochemistry --- DNA (deoxyribonucleic acid) --- biochemie --- PCR (polymerase chain reaction) --- genetische informatie --- Molecular biology --- Analytical biochemistry --- biologie --- moleculaire biologie --- polymeren --- Macromolecules --- Gene Amplification --- Réaction en chaîne de la polymérase --- Technique des traceurs --- Tracer techniques --- État physique --- physical states --- Polymerase Chain Reaction. --- -Chain reaction, Polymerase --- PCR (Biochemistry) --- DNA polymerases --- Amplification, Gene --- Anchored Polymerase Chain Reaction --- Inverse Polymerase Chain Reaction --- Nested Polymerase Chain Reaction --- Anchored PCR --- Inverse PCR --- Nested PCR --- PCR, Anchored --- PCR, Inverse --- PCR, Nested --- Polymerase Chain Reactions --- Reaction, Polymerase Chain --- Reactions, Polymerase Chain --- Ligase Chain Reaction --- -Molecular bases of life. Molecular biology --- -577.2 Molecular bases of life. Molecular biology --- Chain reaction, Polymerase --- Réaction en chaîne de la polymérase --- Polymerase chain reaction --- DNA Replication --- Polymerase Chain Reaction --- 577.2 --- 577.2 Molecular bases of life. Molecular biology --- Molecular bases of life. Molecular biology --- PCR --- Replication, Autonomous --- Autonomous Replication --- Autonomous Replications --- DNA Replications --- Replication, DNA --- Replications, Autonomous --- Replications, DNA --- DNA-Binding Proteins --- Virus Replication --- S Phase --- Laboratory manuals --- #WSCH:WBIO --- Polymerization --- methods --- Laboratory manuals. --- Polymerase chain reaction. --- Dna replication. --- Gene amplification. --- DNA Replication. --- Gene Amplification. --- klonen --- enzymen --- Polymerase chain reaction - Laboratory manuals. --- Genes