Mutation detection : a practical approach

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

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 mu...

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Bibliographic Details
Other Authors: Cotton, Richard G. H., Edkins, E. (Edward), Forrest, S. (Sue)
Format: eBook
Language:English
Published: Oxford ; New York : IRL Press at Oxford University Press, ©1998.
Series:Practical approach series ; 188.
Subjects:
Mutation (Biology) > Laboratory manuals.
Molecular genetics > Laboratory manuals.
Chromosome abnormalities > Laboratory manuals.
Mutation (Biology)
Chromosome abnormalities.
SCIENCE > Life Sciences > Genetics & Genomics.
Chromosome abnormalities
Molecular genetics
Genmutation
Nachweis
Laboratory manuals
Laboratory manuals.
Manuels de laboratoire.
Online Access:Click for online access
Table of 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.

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