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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.
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
Capillary zone electrophoresis : basic concepts -- Capillary zone electrophoresis : methods development -- Capillary zone electrophoresis : secondary equilibrium, micelles, cyclodextrins, and related reagents -- Capillary isoelectric focusing -- Size separations in capillary gels and polymer networks -- Capillary electrochromatography -- Injection -- Detection -- Putting it all together.
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.
In the 1980s, capillary electrophoresis (CE) joined high-performance liquid chromatography (HPLC) as the most powerful separation technique available to analytical chemists and biochemists. Published research using CE grew from 48 papers in the year of commercial introduction (1988) to 1200 in 1997. While only a dozen major pharmaceutical and biotech companies have reduced CE to routine practice, the applications market is showing real or potential growth in key areas, particularly in the DNA marketplace for genomic mapping and forensic identification. For drug development involving small molecules (including chiral separations), one CE instrument can replace 10 liquid chromatographs in terms of speed of analysis. CE also uses aqueous rather than organic solvents and is thus environmentally friendlier than HPLC. The second edition of Practical Capillary Electrophoresis has been extensively reorganized and rewritten to reflect modern usage in the field, with an emphasis on commercially available apparatus and reagents. This authoritative and very comprehensible treatment builds on the author's extensive experience as an instructor of short courses for the American Chemical Society and for industry. - Illustrated with detailed diagrams of electrophoretic phenomena - Offers step-by-step methods development schemes - Presents techniques for developing quantitative, robust, and precise methods - Includes an extensive troubleshooting guide - Updates and greatly expands on the first edition-more than 50% of the text is new - Written by an internationally recognized scientist who is an instructor for American Chemical Society short courses on HPCE
This book presents a selection of current capillary electrophoresis methods used to separate representative types of molecules and particles and in combination with different detection techniques. It includes practical details which are hard to find elsewhere. The volume is intended for beginners in the field and provides an overview of the technique and a starting point for the exploration of the defined literature on different application topics.
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.
Explores the benefits and limitations of the latest capillary electrophoresis techniques Capillary electrophoresis and microchip capillary electrophoresis are powerful analytical tools that are particularly suited for separating and analyzing biomolecules. In comparison with traditional analytical techniques, capillary electrophoresis and microchip capillary electrophoresis offer the benefits of speed, small sample and solvent consumption, low cost, and the possibility of miniaturization. With contributions from a team of leading analytical scientists, Capillary Electrophoresis and Microchip Capillary Electrophoresis explains how researchers can take full advantage of all the latest techniques, emphasizing applications in which capillary electrophoresis has proven superiority over other analytical approaches. The authors not only explore the benefits of each technique, but also the limitations, enabling readers to choose the most appropriate technique to analyze a particular sample. The book's twenty-one chapters explore fundamental aspects of electrophoretically driven separations, instrumentation, sampling techniques, separation modes, detection systems, optimization strategies for method development, and applications. Specific topics include: Critical evaluation of the use of surfactants in capillary electrophoresis Sampling and quantitative analysis in capillary electrophoresis Capillary electrophoresis with electrochemical detection Overcoming challenges in using microchip electrophoresis for extended monitoring applications Capillary electrophoresis of intact unfractionated heparin and related impurities Microchip capillary electrophoresis for in situ planetary exploration Each chapter begins with an introduction and ends with conclusions as well as references to the primary literature. Novices to the field will find this book an easy-to-follow introduction to core capillary electrophoresis techniques and methods. More experienced investigators can turn to the book for troubleshooting tips and expert advice to guide them through the most advanced applications.
Annotation In this volume expert researchers in the field detail the operations of microchip capillary electrophoresis. Chapters focus on small molecule, biomolecule applications, various detection modes, and sample preparation approaches are described. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.Authoritative and practical, Microchip Capillary Electrophoresis Protocol aids scientists in continuing to study microchip capillary electrophoresis.
John R. Petersen and Amin A. Mohammad, along with a panel of leading basic and clinical investigators, review those CE methods that are now replacing many routine serum and blood tests in clinical and forensic laboratories. Major areas reviewed include the coating of columns; the analysis of serum, urine, and CSF proteins and paraproteins; abnormal hemoglobins and hemoglobin Alc; peptides, amino and organic acids; therapeutic drugs; drugs of abuse; viral load; and short tandem repeats (STR). The methods discussed include capillary zone, micellar, electrokinetic, capillary gel, and non-aqueous electrophoresis. Innovative and highly practical, Clinical and Forensic Applications of Capillary Electrophoresis demonstrates the power and versatility of CE-not only to develop new assays, but also to markedly simplify today's clinical and forensic laboratory methodology.