Title Mutation detection : a practical approach / edited by R.G.H. Cotton, E. Edkins, and S. Forrest

Published Oxford ; New York : IRL Press at Oxford University Press, ©1998

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Description 1 online resource (xx, 242 pages) : illustrations Series The practical approach series ; 188 Practical approach series ; 188. Contents Cover -- Contents -- List of Contributors -- Abbreviations -- Introduction -- References -- 1. Single-strand conformation polymorphism analysis -- 1. Introduction -- 2. PCR-SSCP using polyacrylamide slab gel -- PCR Optimization and primer design -- Pre-amplification and isolation by agarose gel electrophoresis -- PCR using [[Sup(32)]P]deoxynucleotide triphosphate -- Removal of 3' appendage -- SSCP gel electrophoresis -- Interpretation of autoradiogram -- Re-amplification and direct sequencing -- Gel matrices other than polyacrylamide Restriction endonuclease fingerprinting and dideoxy fingerprinting -- 3. Fluorescent SSCP in an automated DNA sequencer -- Primer design in post-PCR fluorescent labelling -- Fluorescent labelling by 3' exchange reaction -- SSCP in capillary electrophoresis (CE-SSCP) -- Data processing -- Acknowledgements -- References -- 2. Single-stranded conformation polymorphism and heteroduplex analysis -- 1. Introduction -- 2. Optimization of the PCR reaction -- 3. SSCP sample prepration -- 4. Optimization of SSCP/HA detection -- 5. Multiplexing -- 6. Interpretation of results -- 7. Applications 8. Other methods -- References -- 3. Comprehensive mutation detection with denaturing gradient gel electrophoresis -- 1. Introduction -- The scope of DGGE, its distinctive capabilities, and the nature of results -- 2. Background -- 3. Basic principle, the physical properties of DNA -- 4. Overview of the procedures in searching for mutants -- Defining segments for scrutiny -- Sample preparation -- Gradient and velocity separations -- Features of the gel patterns -- Discrimination of zygozygosity -- Comments -- 5. Use of the psoralen cross-link as a clamp -- The psoralen protocol 6. Computational tools -- What is a meltmap? -- Meltmap protocol -- Predicting electrophoretic separations -- Computer operations for MUTRAV -- 7. Other members of the DGGE family -- Gel separations in a uniform, partially denaturing environment -- Capillary electrophoresis -- The thermal gradient -- The temperature ramp -- 2D length and gradient separations -- 8. End notes -- Acknowledgments -- References -- 4. Cleavage using RNase to detect mutations -- 1. Introduction -- 2. RNase protection assay for mutation detection -- Evaluation of the sensitivity -- Source material PCR for RNase protection assay -- RNA probe preparation -- RNase protection -- Detection of digested probe -- Mutation detection by sequencing of the PCR products -- Other modified methodologies for mutation detection -- Acknowledgements -- References -- 5. Cleavage of mismatched bases using chemical reagents -- 1. Introduction -- 2. Basic procedures -- Comments on the basic procedures -- 3. Ultra fast chemical mismatch detection -- Labelling -- Solid phase -- Comments -- References -- 6. Mutation detection using T4 endonuclease VII -- 1. Introduction -- 2. The biology of Endo VII Summary Mutation detection is increasingly undertaken in a wide spectrum of research areas: in medicine it is fundamental in isolating disease genes and diagnosis, and is especially important in cancer research; in biology, commercially important genes can be identified by the mutations they contain. But mutation detection is time-consuming and expensive. This volume offers tried and tested protocols for a range of detection methods, from the labs of researchers in the field Bibliography Includes bibliographical references and index Notes Master and use copy. Digital master created according to Benchmark for Faithful Digital Reproductions of Monographs and Serials, Version 1. Digital Library Federation, December 2002. http://purl.oclc.org/DLF/benchrepro0212 MiAaHDL digitized 2010 HathiTrust Digital Library committed to preserve pda MiAaHDL Print version record Subject Mutation (Biology) -- Laboratory manuals Molecular genetics -- Laboratory manuals Chromosome abnormalities -- Laboratory manuals Mutation (Biology) Chromosome abnormalities. Mutation Molecular Biology Chromosome Aberrations SCIENCE -- Life Sciences -- Genetics & Genomics. Chromosome abnormalities Molecular genetics Mutation (Biology) Genmutation Nachweis Genre/Form Laboratory manuals Laboratory manuals. Manuels de laboratoire. Form Electronic book Author Cotton, Richard G. H Edkins, E. (Edward) Forrest, S. (Sue) ISBN 0585484139 9780585484136 1280375485 9781280375484 9780191565694 0191565695

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