PCR Mutation Detection Protocols

Bimal D. Theophilus and Ralph Rapley provide biological and clinical investigators with a comprehensive collection of new, recent, and updated PCR-based screening methods suitable for detecting the presence of both known and novel mutations. The methods cover point mutations (e.g., ASO-PCR, SSCP, DGGE, chemical cleavage), deletions (multiplex PCR, FISH, blotting), non-sense mutations (PTT), and more. The new and exciting techniques of DNA array analysis, along with such recently developed experimental methods as conformation-sensitive gel electrophoresis, are also included. Each chapter explains the basic theory behind the technique and provides valuable notes essential for its successful execution.

Table of Contents:
Agarose and Polyacrylamide Gel Electrophoresis Andrea M. Guilliatt Internal Labeling of DNA Probes Ralph Rapley and Bimal D. M. Theophilus End-Labeling of DNA Probes Adrian J. Harwood Southern Blotting of Agarose Gels by Capillary Transfer Ralph Rapley and Ian J. Williams Restriction Fragment Length Polymorphism Mohammad S. Enayat PCR: Principles, Procedures, and Parameters Nicola Louise Jones Allele-Specific Oligonucleotide PCR Elaine K. Green Long-Range PCR, Peter A. Davies and George Gray Analysis of Nucleotide Sequence Variations by Solid-Phase Minisequencing Anu Wartiovaara and Ann-Christine Syvanen Cycle Sequencing of PCR Products G. K. Surdhar Fluorescent In Situ Hybridization Sara A. Dyer and Elaine K. Green. The Protein Truncation Test Carol A. Hardy Mutation Detection in Factor VIII cDNA from Lymphocytes of Haemophilia A Patients by Solid Phase Fluorescent Chemical Cleavage of Mismatch Naushin H. Waseem, Richard Bagnall, Peter M. Green, and Francesco Giannelli Denaturing Gradient Gel Electrophoresis Yvonne Wallis Conformation-Sensitive Gel Electrophoresis Ian J. Williams and Anne C. Goodeve SSCP/Heteroduplex Analysis Andrew J. Wallace Cleavase(R) Fragment Length Polymorphism Analysis for Genotyping and Mutation Detection Laura Heisler and Chao-Hung Lee Automated Genotyping Using the DNA MassArraya Technology Christian Jurinke, Dirk van den Boom, Charles R. Cantor, and Hubert Koster An Introduction to Bioinformatics Henry Brzeski