Capillary Electrophoresis of Nucleic Acids

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Januar 2001



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 to utilize this new resource for analysis of genetic variation and for the detection of disease causing mutations. The advent of capillary electrophoresis (CE), with its power to separate and analyze 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 analysis are manifold. Further, the high sensitivity of detection, and the ab- ity to increase sample throughput with parallel analysis, has led to the creation of a full range of analysis of DNA molecules, from modified DNA-adducts and single-strand oligonucleotides through to PCR-amplified DNA fragments and whole chromosomes. Capillary Electrophoresis of Nucleic Acids focuses on such analytical protocols, which can be used for detection and analysis of mutations and modification, from precise DNA loci through to 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.


Part I. Practical Applications for Rapid DNA Fragment Sizing and Analysis. Development of a High-Throughput Capillary Electrophoresis Protocol for DNA Fragment Analysis, H. Michael Wenz, David Dailey, and Martin D. Johnson. Ultra-Fast DNA Separations Using Capillary Electrophoresis, Karel Klepárník, Odilo M. Mueller, and Frantisek Foret. Fast DNA Fragment Sizing in a Short Capillary Column, Haleem J. Issaq and King C. Chan.
Part II. Practical Applications for Mutation Detection. Detection of DNA Polymorphisms Using PCR-RFLP and Capillary Electrophoresis, John M. Butler and Dennis J. Reeder. High Resolution Analysis of Point Mutations by Constant Denaturant Capillary Electrophoresis(CDCE), Konstantin Khrapko, Hilary
A. Coller, Xiao-Cheng Li-Sucholeiki, Paulo C. André, and William G. Thilly. Point Mutation Detection by Temperature-Programmed Capillary Electrophoresis, Cecilia Gelfi, Laura Cremoresi, Maurizio Ferrari, and Pier Giorgio Righetti. Single-Nucleotide Primer Extension Assay by Capillary Electrophoresis Laser-Induced Fluorescence, Christine A. Piggee and Barry L. Karger. Amplification Refractory Mutation System Analysis of Point Mutations by Capillary Electrophoresis, Paola Carrera, Pier Giorgio Righetti, Cecilia Gelfi, and Maurizio Ferrari. SSCP Analysis of Point Mutations by Multicolor Capillary Electrophoresis, Kenshi Hayashi, H. Michael Wenz, Masakazu Inazuka, Tomoko Tahira, Tomonari Sasaki, and Donald H. Atha. SSCP Analysis by Capillary Electrophoresis with Laser-Induced Fluorescence Detector, Jicun Ren. Magnetic Bead-Isolated Single-Strand DNA for SSCP Analysis, Takao Kasuga, Jing Cheng, and Keith R. Mitchelson.
Part III. Practical Applications for Genetic Analysis. Analysis of Short Tandem Repeat Markers by Capillary Array Electrophoresis, Elaine S. Mansfield, Robert
B. Wilson, and Paolo Fortina. Genotyping by Microdevice Electrophoresis, Dieter Schmalzing, Lance Koutny, Aram Adourian, Dan Chisholm, Paul Matsudaira, and Daniel Ehrlich. Applicationsof Constant Denaturant Capillary Electrophoresis and Complimentary Procedures: Measurement of Point Mutational Spectra, Andrea S. Kim, Xiao-Cheng Li-Sucholeiki, and William G. Thilly. Toward Effective PCR-Based Amplification of DNA on Microfabricated Chips, Jerome P. Ferrance, Braden Giordano, and James P. Landers. Low-Stringency Single Specific Primer-Amplified DNA Analysis by Capillary Electrophoresis, Michael A. Marino. DOP-PCR Amplification of Whole Genomic DNA and Microchip-Based Capillary Electrophoresis, Paolo Fortina, Jing Cheng, Larry J. Kricka, Larry
C. Waters, Stephen C. Jacobson, Peter Wilding, and J. Michael Ramsey. Analysis of Triplet-Repeat DNA by Capillary Electrophoresis, Yuriko Kiba and Yoshinobu Baba. Chemical Mismatch Cleavage Analysis by Capillary Electrophoresis with Laser-Induced Fluorescence Detection, Jicun Ren.
Part IV. Practical Applications for Analysis of RNA and Gene Expression. Quantitiation of mRNA by Competitive PCR Using Capillary Electrophoresis, Stephen J. Williams and P. Mickey Williams. Quantitative RT-PCR from Fixed Paraffin-Embedded Tissues by Capillary Electrophoresis, Giorgio Stanta, Serena Bonin, and Maurizio Lugli. Differential Display Analysis by Capillary Electrophoresis, Xilin Zhao and Kimberly S. George.
Part V. Practical Applications for DNA Sequence Analysis. Capillary Array Electrophoresis Analyzer, Masao Kamahori and Hideki Kambara. DNA Sequencing at Elevated Temperature by Capillary Electrophoresis, Peter Lindberg and Johan Roeraade. DNA Sequencing in Noncovalently Coated Capillaries Using Low Viscosity Polymer Solutions, Ramakrishna Madabhushi. Selective Primer Sequencing from a DNA Mixture by Capillary Electrophoresis, Tao Li, Kazunori Okano, and Hideki Kambara. Sequencing of RAPD Fragments Using 3'-Extended Oligonucleotide Primers, John Davis, Ivan Biros, Ian Findlay, and Keith Mitchelson.
Part VI. Applications for Analysis of DNA-Protein and DNA-Ligand Interactions. Capillary Affinity Gel Electrophoresis, Yos


"These volumes comprehensively and accurately present CE technology and techniques with an emphasis on providing an understanding of the theory and instrumentation (Vol. 1), as well as detailed examples of successful analytical protocols (both volumes)....Chapters on the use of microchip and mass spectrometric detection are also included....Although numerous experimental protocols can be found in Vol. 1, it is clear that the intent is to communicate the underlying theory and capability of CE for separations. Vol. 1 should be of interest to the novice as well as the expert. In Vol. 2, each of the 31 chapters present specific examples of the use of CE. ...Nonetheless, these two volumes of the Methods in Molecular Biology series offer a representative and needed overview of the current theory and practice of CE of nucleic acids. These texts should prove very useful to both active investigators in this area and those seeking to learn more about the use and and capability of CE." - Clinical Chemistry
EAN: 9780896037656
ISBN: 0896037657
Untertitel: 2001. Auflage. Book. Sprache: Englisch.
Verlag: Humana Press
Erscheinungsdatum: Januar 2001
Seitenanzahl: 432 Seiten
Format: gebunden
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