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Full Description
The newest and most up-to-date collection, PCR Protocols offers detailed laboratory procedures for the use of polymerase chain reactions in a wide range of applications. Although specific examples and experimental systems are described, the book's focus is on general applications that can be modified to suit a broad spectrum of research systems, thus making its scope appealing to nearly all researchers. PCR Protocols features clear, easy-to-follow descriptions of procedures and a Notes section in each chapter to provide tips, alternative suggestions, and other enhancements of the protocols. Key Topics and Techniques: basic startup procedures for PCR PCR in detecting DNA and RNA PCR in DNA synthesis and mutagenesis PCR cloning of DNA selection of primers radioactive and nonradioactive labeling quantitation of PCR products by HPLC chromosome assignment PCR mapping of HLA genes sequencing of PCR products genomic footprinting quantitation of tumor gene expression screening of phage libraries SSP-PCR and genome walking subcloning of PCR products.
Contents
Polymerase Chain Reaction. Selection of Primers for Polymerase Chain Reaction. Direct Radioactive Labeling of Polymerase Chain Reaction Products. Use of Arithmetic Polymerase Chain Reaction for Synthesis of Single-Stranded Probes for S1 Nuclease Assays. Nonradioactive Labeling of Polymerase Chain Reaction Products. Quantitation and Purification of Polymerase Chain Reaction Products by High-Performance Liquid Chromatography. Use of Polymerase Chain Reaction for Screening Transgenic Mice. Polymerase Chain Reaction Analysis of DNA from Paraffin-Embedded Tissue. The Use of Polymerase Chain Reaction for Chromosome Assignment. Mapping MHC Class II Genes and Disease-Susceptibility. The Use of the Polymerase Chain Reaction and the Detection of Amplified Products. Determination of Loss of Heterozygosity Using Polymerase Chain Reaction. Direct Sequencing of Polymerase Chain Reaction Products. Manual and Automated Direct Sequencing of Product Generated by the Polymerase Chain Reaction. Genomic Footprinting by Ligation Mediated Polymerase Chain Reaction. RNA Template-Specific Polymerase Chain Reaction (RS-PCR). Quantitative Measurement of Relative Gene Expression in Human Tumors. Identification of Alternatively Spliced mRNAs and Localization of 5' Ends by Polymerase Chain Reaction Amplification. Utilization of Polymerase Chain Reaction for Clonal Analysis of Gene Expression. Sequencing DNA Amplified Directly from a Bacterial Colony. Use of Polymerase Chain Reaction to Screen Phage Libraries. Molecular Cloning of Polymerase Chain Reaction Fragments with Cohesive Ends. Rapid (Ligase-Free) Subcloning of Polymerase Chain Reaction Products. Use of Polymerase Chain Reaction for Making Recombinant Constructs. In Vitro Recombination and Mutagenesis of DNA. Use of Polymerase Chain Reaction for the Rapid Construction of Synthetic Genes. Recombinant Circle Polymerase Chain Reaction for Site-Directed Mutagenesis. Site-Directed Mutagenesis by Double Polymerase Chain Reaction. Generation of a Polymerase Chain Reaction Renewable Source of Subtractive cDNA. PCR-Based Full-Length cDNA Cloning Utilizing the Universal-Adaptor/Specific DOS Primer-Pair Strategy. Use of Degenerate Oligonucleotide Primers and the Polymerase Chain Reaction to Clone Gene Family Members. Single Specific Primer-Polymerase Chain Reaction (SSP-PCR) and Genome Walking. cDNA Cloning by Inverse Polymerase Chain Reaction. Amplification of Gene Ends from Gene Libraries by Polymerase Chain Reaction with Single-Sided Specificity. Anchoring a Defined Sequence to the 5' Ends of mRNAs. Index.
