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Recent advances in the biosciences have led to a range of powerful new technologies, particularly nucleic acid, protein and cell-based methodologies. The most recent insights have come to affect how scientists investigate and define cellular processes at the molecular level. Molecular Biomethods Handbook, 2nd Edition expands upon the techniques included in the first edition, providing theory, outlines of practical procedures, and applications for a range of techniques. Part A of the book describes nucleic acid methods, such as gene expression profiling, microarray analysis and quantitative PCR. In Part B, protein and cell-based methods are outlined, in subjects ranging from protein engineering to high throughput screening. Written by a well-established panel of research scientists, Molecular Biomethods Handbook, 2nd Edition provides an up to date collection of methods used regularly in the authors' own research programs. This book will prove to be an invaluable reference for those engaged in or entering the field of molecular biology, and will provide the necessary background for those interested in setting up and using the latest molecular techniques.
Comprehensive collection of wide-ranging molecular and biochemical techniques frequently used in medical and clinical research laboratories around the world Covers a wide array of techniques and protocols used for diagnostics Includes step-by-step reproducible laboratory protocols written by experts in the field and includes a notes section that outlines tricks of the trade Provides in-depth understanding of the theory, practice and application of each technique
Contenu
Nucleic Acid Methods.- The Manipulation of Nucleic Acids.- Restriction Enzymes.- Principles and Medical Applications of the Polymerase Chain Reaction.- Probe Design, Production, and Applications.- Southern Blotting as a Diagnostic Method.- Capillary Electrophoresis of DNA.- Denaturing High-Performance Liquid Chromatography (DHPLC) for Nucleic Acid Analysis.- Denaturing Gradient Gel Electrophoresis (DGGE).- Single Strand Conformation Polymorphism (SSCP) Analysis.- Randomly Amplified Polymorphic DNA (RAPD).- Quantification of mRNA Using Real Time RT-PCR.- Quantitative Analysis of DNA Sequences by PCR and Solid-Phase Minisequencing.- Multiplex Amplifiable Probe Hybridization (MAPH).- Gene Expression Profiling.- Comparative Genomic Hybridization in Clinical and Medical Research.- Subtractive Hybridization.- Fluorescence In Situ Hybridization.- Quantitative Trait Locus Mapping to Identify Genes for Complex Traits in Mice.- cDNA Microarrays.- Mapping Techniques.- Single Nucleotide Polymorphisms.- Gene/Protein Sequence Analysis.- Protein and Cell Methods.- Protein Electrophoresis.- Protein Blotting.- Capillary Electrophoresis of Proteins.- Autoradiography and Fluorography.- Mass Spectrometry of Proteins and Peptides.- Post-Translational Modif ications of Proteins.- Protein Microarray Technology.- ProteinProtein Interactions.- Glycoprotein Analysis.- Solid-Phase Peptide Synthesis.- Monoclonal Antibodies.- Antibody Phage Display.- Protein Engineering.- Directed Protein Evolution.- Enzyme Linked Immunosorbent Assay (ELISA).- Epitope Mapping.- Quantum Dots.- Ion-Exchange Chromatography.- Size-Exclusion Chromatography.- Hydrophobic Interaction Chromatography.- Affinity Chromatography.- High Performance Liquid Chromatography (HPLC) of Peptides and Proteins.- Amino Acid Analysis.- Surface Plasmon Resonance.- Macromolecular Crystallography.- Microchip Devices for Bioanalysis.- Mammalian Cell Culture.- Plant Tissue Culture.- Stem Cells and Regenerative Medicine.- Cryopreservation.- Magnetic Resonance Imaging.- Electron Microscopy.- Confocal Microscopy.- Laser Microdissection.- Biomedical Uses of Flow Cytometry.- Immunomicroscopy.- In Situ Hybridization.- High Throughput Screening.