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Mutation detection : a practical approach /
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...
| Clasificación: | Libro Electrónico |
|---|---|
| Otros Autores: | , , |
| Formato: | Electrónico eBook |
| Idioma: | Inglés |
| Publicado: |
Oxford ; New York :
IRL Press at Oxford University Press,
©1998.
|
| Colección: | Practical approach series ;
188. |
| Temas: | |
| Acceso en línea: | Texto completo |
Tabla de Contenidos:
- 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.