Capillary Electrophoresis of Nucleic Acids

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.

Includes supplementary material: sn.pub/extras

Klappentext
The advent of capillary electrophoresis (CE) , with its power to separate and analyze very small amounts of DNA, has generated a host of DNA analytical procedures. In these novel and wide-ranging volumes of Capillary Electrophoresis of Nucleic Acids, an outstanding panel of hands-on experts and developers of CE equipment describe in step-by-step fashion their best cutting-edge methods for the detection and analysis of DNA mutations and modifications, ranging from precise DNA loci to entire genomes of organisms. This first volume of the set, Introduction to the Capillary Electrophoresis of Nucleic Acids, covers the practical and theoretical considerations behind the use of capillary electrophoresis for the analysis of small oligonucleotides and modified nucleotides. Along with detailed instructions ensuring ready reproducibility, these protocols offer time-tested advice on instrumentation, signal detection, the capillary environment, and the integration of mass spectrometry with CE. Several chapters are devoted to the analysis of small therapeutic oligonucleotides, nucleosides, and ribonucleotides by CE. The companion volume, Practical Applications of Capillary Electrophoresis, addresses techniques for high-throughput analysis of DNA fragments using SNP detection, mutation detection, DNA sequencing methods, and DNA-ligand interactions.
Comprehensive and up-to-date, the paired volumes of Capillary Electrophoresis of Nucleic Acids offer an authoritative guide with easy access to fast, versatile, reliable, and powerful technologies for all those basic and clinical investigators analyzing DNA variation today.

Inhalt
Analytical Considerations for Capillary Electrophoresis.- Overview: The Application of Capillary Electrophoresis for DNA Polymorphism Analysis.- DNA Separation Mechanisms During Electrophoresis.- Purification Methods for Preparing Polymerase Chain Reaction Products for Capillary Electrophoresis Analysis.- Quantitative Measurements.- Microchip-Based Capillary Electrophoresis Systems.- DNA Sequencing by Capillary Array Electrophoresis.- 7 Determining Dye-Induced DNA Mobility Shifts for DNA Sequencing Fragments by Capillary Electrophoresis.- Capillary Matrix: Considerations for Capillary Electrophoresis.- Influence of Polymer Concentration and Polymer Composition on Capillary Electrophoresis of DNA.- Capillary Coatings.- Replaceable Polymers for DNA Sequencing by Capillary Electrophoresis.- Sieving Matrix Selection.- Parameters Affecting Capillary Electrophoretic Separation of DNA.- Electrophoresis Matrix: Applications for DNA Fragment Sizing.- DNA Separation by Capillary Electrophoresis in Lyotropic Polymer Liquid Crystals.- Capillary Electrophoresis of DNA Fragments Using Poly(Ethylene Oxide) as a Sieving Material.- DNA Capillary Electrophoresis Using Block Copolymer as a New Separation Medium.- 16 DNA Analysis Under Highly Denaturing Conditions in Bare Fused Silica Capillaries.- Capillary Electrophoresis with Glycerol as an Additive.- Capillary Electrophoresis of DNA Fragments with Replaceable Low-Gelling Agarose Gels.- Practical Applications for Large DNA Fragment Sizing and Fragment Collection.- Robust Field Inversion Capillary Electrophoretic Separation of Long DNA Fragments.- Pulsed-Field Capillary Electrophoresis Separation of Large DNA Fragments.- Collection of Capillary Electrophoresis Fractions on a Moving Membrane.- Separation of Supercoiled DNA Using CapillaryElectrophoresis.- Practical Applications for Oligonucleotide and Nucleotide Analysis.- Quality Control of Nucleotides and Primers for PCR.- Analysis of Modified Oligonucleotides with Capillary Gel Electrophoresis.- Use of Capillary Electrophoresis for Concentration Analysis of Phosphorothioate Oligonucleotides.- Capillary Electrophoresis Separation of Ribonucleosides.- On-Line Preconcentration and Separation of Antisense Oligonucleotides by ITP-CE in Dextran-Filled Capillaries.- Practical Applications for Analysis of Environmental Damage and Cellular Metabolism of DNA.- Capillary Electrophoresis-Mass Spectrometric Analysis of DNA Adducts.- Analysis of Environment-Induced DNA Damage by Capillary Electrophoresis.- Assay for DNA Damage Using Immunochemical Recognition and Capillary Electrophoresis.- Analysis of DNA Damage Using Capillary Zone Electrophoresis and Electrospray Mass Spectrometry.- Integration of Phosphodiesterase-Induced Degradation of Oligonucleotides with Capillary Polymer-Sieving Electrophoresis.- Capillary Electrophoresis of Urinary Normal and Modified Nucleosides of Cancer Patients.