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The development of PCR, which enables extremely small amounts of DNA to be amplified, led to the rapid development of a multiplicity of a- lytical procedures that permit use of this new resource for the analysis of genetic variation and for the detection of disease-causing mutations. The advent of capillary electrophoresis (CE), with its power to separate and a- lyze very small amounts of DNA, has also stimulated researchers to develop analytical procedures for the CE format. The advantages of CE in terms of speed and reproducibility of analyses are manifold. Furthermore, the high s- sitivity of detection, and the ability to increase sample throughput with par- lel analysis, has led to the creation of a full range of analysis of DNA molecules, from modified DNA adducts and single-strand oligonucleotides through PCR-amplified DNA fragments and whole chromosomes. Capillary Elect- phoresis of Nucleic Acids focuses on analytical protocols that can be used for detection and analysis of mutations and modification, from precise DNA loci through entire genomes of organisms. Important practical considerations for CE, such as the choice of separation media, electrophoresis conditions, and the influence of buffer additives and dyes on DNA mobility, are discussed in several key chapters and within particular applications.
Die Kapillarelektrophorese (CE) ist heute in biochemischen Laboratorien noch nicht als Routinemethode zur DNA-Trennung etabliert, obwohl sie diverse Vorzüge gegenüber den derzeitigen Methoden besitzt. In diesem Band stellen verschiedene Wissenschaftler die theoretischen Aspekte der Trennung von Oligonucleinsäuren mit der CE, die instrumentellen Grundlagen und deren Grenzen und schließlich die praktische Anwendung in der Biochemie und Molekularbiologie vor. This book on capillary electrophoresis is unique in its focus on the separation of nucleic acids. The importance of electrophoretic separation in every molecular biology laboratory justifies this specialization, which is also reflected in the development of instrumentation. This book is aimed to help to implement this rather new and promising technology in the biological laboratories and to help to overcome typical problems that can occur when starting with a new technique. It should also trigger further development in the field. It covers the theoretical background as well as practical examples of usual applications. The authors are all experts in their field, having many years of experience with capillary electrophoresis and their application to nucleic acids.
Capillary Gel Electrophoresis and Related Microseparation Techniques covers all theoretical and practical aspects of capillary gel electrophoresis. It also provides an excellent overview of the key application areas of nucleic acid, protein and complex carbohydrate analysis, affinity-based methodologies, micropreparative aspects and related microseparation methods. It not only gives readers a better understanding of how to utilize this technology, but also provides insights into how to determine which method will provide the best technical solutions to particular problems. This book can also serve as a textbook for undergraduate and graduate courses in analytical chemistry, analytical biochemistry, molecular biology and biotechnology courses. Covers all theoretical and practical aspects of capillary gel electrophoresis Excellent overview of the key applications of nucleic acid, protein and complex carbohydrate analysis, affinity-based methodologies, micropreparative aspects and related microseparation methods Teaches readers how to use the technology and select methods that are ideal for fundamental problems Can serve as a textbook for undergraduate and graduate courses in analytical chemistry, analytical biochemistry, molecular biology and biotechnology courses
The development of PCR, which enables extremely small amounts of DNA to be amplified, led to the rapid development of a multiplicity of a- lytical procedures that permit use of this new resource for the analysis of genetic variation and for the detection of disease-causing mutations. The advent of capillary electrophoresis (CE), with its power to separate and a- lyze very small amounts of DNA, has also stimulated researchers to develop analytical procedures for the CE format. The advantages of CE in terms of speed and reproducibility of analyses are manifold. Furthermore, the high s- sitivity of detection, and the ability to increase sample throughput with par- lel analysis, has led to the creation of a full range of analysis of DNA molecules, from modified DNA adducts and single-strand oligonucleotides through PCR-amplified DNA fragments and whole chromosomes. Capillary Elect- phoresis of Nucleic Acids focuses on analytical protocols that can be used for detection and analysis of mutations and modification, from precise DNA loci through entire genomes of organisms. Important practical considerations for CE, such as the choice of separation media, electrophoresis conditions, and the influence of buffer additives and dyes on DNA mobility, are discussed in several key chapters and within particular applications.
Various sophisticated techniques such as capillary electrophoresis, pulsed-field electrophoresis, fingerprinting using RFLP and RAPD, DNA sequencing, and mobility shift assay are described here in detail. Leading experts present the required apparatus, appropriate use, preparation of probes, gel staining, interpretation of results, tricks for troubleshooting, manufacturers' addresses, helpful Internet resources, as well as specific applications, e.g. in legal medicine, microbiology and agriculture.
In Clinical Applications of Capillary Electrophoresis, Stephen Palfrey brings together for first time a collection of detailed capillary electrophoresis protocols designed exclusively for clinical applications. Written by the leading scientists who have often perfected these methods in their own laboratories, the protocols furnish new and more powerful assays for many routine serum and blood tests now regularly performed in clinical laboratories, including urine protein analysis, hemoglobin separation, and the detection of CSF proteins, lipoproteins, myoglobin, cryoglobulins, HbA1c, and cathepsin. The protocols offered for DNA studies include double-stranded DNA analysis, the prenatal diagnosis of Down's syndrome, Rh D/d genotyping, the identification of mutated p53 oncogene, and the detection of microsatellite instability in cancers. Many of the methods can be automated to replace the more costly and labor-intensive tests that are currently used in most clinical laboratories. Clinical Applications of Capillary Electrophoresis demonstrates clearly the simplicity, versatility, and power of CE over conventional methods. It offers to beginning clinical investigators, as well as established laboratories new to the technique, a representative range of highly practical CE methods-assays that are not only certain to become ever more productive, but are already eminently useful today.