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 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 singlestrand 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.

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 produced a host of DNA analytical procedures. In the
Unprecedentedly 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 second volume of the set, Practical Applications of Capillary Electrophoresis, covers techniques for high-throughput analysis of DNA fragments using SNP detection, mutation detection, DNA sequencing methods, and DNA-ligand interactions. Several chapters also discuss CE analysis at elevated temperatures and the use of micro-CE and array-CE devices. The companion volume, Introduction to the Capillary Electrophoresis of Nucleic Acids, offers readily reproducible methods for the analysis of small oligonucleotides and modified nucleotides, and time-tested advice on instrumentation, signal detection, the capillary environment, and the integration of mass spectrometry with CE.
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
Practical Applications for Rapid DNA Fragment Sizing and Analysis.- Development of a High-Throughput Capillary Electrophoresis Protocol for DNA Fragment Analysis.- Ultra-Fast DNA Separations Using Capillary Electrophoresis.- Fast DNA Fragment Sizing in a Short Capillary Column.- Practical Applications for Mutation Detection.- Detection of DNA Polymorphisms Using PCR-RFLP and Capillary Electrophoresis.- High Resolution Analysis of Point Mutations by Constant Denaturant Capillary Electrophoresis (CDCE).- Point Mutation Detection by Temperature-Programmed Capillary Electrophoresis.- Single-Nucleotide Primer Extension Assay by Capillary Electrophoresis Laser-Induced Fluorescence.- Amplification Refractory Mutation System Analysis of Point Mutations by Capillary Electrophoresis.- SSCP Analysis of Point Mutations by Multicolor Capillary Electrophoresis.- SSCP Analysis by Capillary Electrophoresis with Laser-Induced Fluorescence Detector.- Magnetic Bead-Isolated Single-Strand DNA for SSCP Analysis.- Practical Applications for Genetic Analysis.- Analysis of Short Tandem Repeat Markers by Capillary Array Electrophoresis.- Genotyping by Microdevice Electrophoresis.- Applications of Constant Denaturant Capillary Electrophoresis and Complementary Procedures.- Toward Effective PCR-Based Amplification of DNA on Microfabricated Chips.- Low-Stringency Single Specific Primer-Amplified DNA Analysis by Capillary Electrophoresis.- DOP-PCR Amplification of Whole Genomic DNA and Microchip-Based Capillary Electrophoresis.- Analysis of Triplet-Repeat DNA by Capillary Electrophoresis.- Chemical Mismatch Cleavage Analysis by Capillary Electrophoresis with Laser-Induced Fluorescence Detection.- Practical Applications for Analysis of RNA and Gene Expression.- Quantitation of mRNA by Competitive PCR Using Capillary Electrophoresis.- Quantitative RT-PCR from Fixed Paraffin-Embedded Tissues by Capillary Electrophoresis.- Differential Display Analysis by Capillary Electrophoresis.- Practical Applications for DNA Sequence Analysis.- Capillary Array Electrophoresis Analyzer.- DNA Sequencing at Elevated Temperature by Capillary Electrophoresis.- DNA Sequencing in Noncovalently Coated Capillaries Using Low Viscosity Polymer Solutions.- Selective Primer Sequencing from a DNA Mixture by Capillary Electrophoresis.- Sequencing of RAPD Fragments Using 3'-Extended Oligonucleotide Primers.- Applications for Analysis of DNA-Protein and DNA-Ligand Interactions.- Capillary Affinity Gel Electrophoresis.- Capillary DNA-Protein Mobility Shift Assay.- Protein-DNA Binding Affinities by Capillary Electrophoresis.- Ligand Binding to Oligonucleotides.