John Walker and Ralph Rapley have collected a wide-ranging group of molecular and biochemical techniques that are the most frequently used in medical and clinical research, especially diagnostics. The authors-well-established investigators who run their own research programs and use the methods on a regular basis-outline the practical procedures for using them and describe a variety of pertinent applications. Among the technologies presented are southern and western blotting, electrophoresis, PCR, c DNA and protein microarrays, liquid chromatography, in situ hybridization, karyotyping, flow cytometry, bioinformatics, genomics, and ribotyping. The applications include assays for mutation detection, m RNA analysis, chromosome translocations, inborn errors of metabolism, protein therapeutics, and gene therapy.
Зміст
Basic Techniques in Molecular Biology.- Probe Design, Production, and Applications.- Restriction Enzymes.- Southern Blotting as a Diagnostic Method.- Western Blotting as a Diagnostic Method.- Principles and Medical Applications of the Polymerase Chain Reaction.- Single-Strand Conformation Polymorphism (SSCP) Analysis.- Denaturing Gradient Gel Electrophoresis (DGGE).- Quantitative Analysis of DNA Sequences by PCR and Solid-Phase Minisequencing.- to Capillary Electrophoresis of DNA.- Mapping Techniques.- Dideoxyfingerprinting for Mutation Detection.- Conformation-Sensitive Gel Electrophoresis.- Amplification Refractory Mutation System and Molecular Diagnostics.- Ligase Chain Reaction.- Chemical Cleavage of Mismatch.- The Protein Truncation Test.- Linkage, Allele Sharing, and Association.- Single-Nucleotide Polymorphisms.- c DNA Microarrays.- Nucleic Acid Sequence-Based Amplification.- Oligonucleotide Ligation Assay.- Quantitative Taq Man Real-Time PCR.- Use of Denaturing High-Performance Liquid Chromatography in Molecular Medicine.- Quantitative PCR.- Liquid Chromatography-Mass Spectrometry of Nucleic Acids.- Comparative Genomic Hybridization in Clinical and Medical Research.- Bioinformatic Tools for Gene and Protein Sequence Analysis.- In Situ Hybridization.- Enzyme-Linked Immunosorbent Assay.- Protein Therapeutics.- Karyotyping.- Microsatellite Analysis.- Analysis of Chromosomal Translocations.- Differential Display.- Techniques for Gene Expression Profiling.- Capillary Electrophoresis in Clinical Analysis.- Flow Cytometry in the Biomedical Arena.- Immunocytochemistry.- Ribotyping in Clinical Microbiology.- Diagnostic Applications of Protein Microarrays.- The Human Genome Project.- Prenatal Diagnosis of Inborn Errors of Metabolism.- Gene Therapy.