Capillary Electrophoresis of Nucleic Acids / Edition 1

by Keith R. Mitchelson, Jing Cheng

Overview

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 prools, 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.

Product Details

ISBN-13:	9780896037656
Publisher:	Springer-Verlag New York, LLC
Publication date:	01/19/2001
Series:	Methods in Molecular Biology , #163
Edition description:	2001
Pages:	408
Product dimensions:	5.98(w) x 9.02(h) x 0.04(d)

About the Author

Mitchelson, Keith R. (Univ of Queensland); Cheng, Jing

Table of Contents

Practical Applications for Rapid DNA Fragment Sizing and Analysis.- Development of a High-Throughput Capillary Electrophoresis Prool 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.

Editorial Reviews

Reviewer: Eugene A Davidson, PhD (Georgetown University School of Medicine)
Description: This volume is a companion to one in which the theory and methods of capillary electrophoresis (CE) are highlighted.
Purpose: The purpose is to show how to use CE to analyze nucleic acids.
Audience: The intended audience is fellows, researchers, and students.
Features: Practical applications of capillary electrophoresis to contemporary problems in nucleic acid analysis are reviewed. For each of the procedures, protocols are presented with sufficient detail as to allow investigators to apply them in the laboratory. Each of the 31 chapters is accompanied by a short, focused bibliography. The volume begins with descriptions of general separations using CE, including methods emphasizing short analysis times. This is followed by procedures for analysis of polymorphisms, PCR products, triplet repeats, and approaches to sequencing. There is a chapter on differential display, an area of considerable interest given the availability of genomic sequences.
Assessment: Techniques such as CE are well-suited to working with small amounts of material and providing rapid, high-resolution results. Hence, this volume will be of value to all investigators requiring analysis of DNA fragments, sequences, and conformation.

Doody's Review Service

"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

From the Publisher

3 Stars from Doody

Explores techniques for high-throughput analysis of DNA fragments using SNP detection, mutation detection, DNA sequencing methods, and DNA-ligand interactions, capillary electrophoresis (CE) analysis at high temperatures and the use of micro-CE and array-CE devices. Articles also cover techniques for quantitative estimation of gene compression using CE, and reviews practical choices for separation media and the CE environment DNA sequencing. Illustrated with diagrams and graphs. Annotation c. Book News, Inc., Portland, OR (booknews.com)

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