Faced with the search for genes associated with human diseases and the prospect of mapping and sequencing the human genome and those of other organisms, scientists must employ new strategies designed to map and clone large regions of DNA. Written by internationally renowned scientists, and containing contributions from some of the leading laboratories in the world, Techniques for the Analysis of Complex Genomes describes genomic mapping by pulsed field gel electrophoresis, and provides a detailed account of the cloning of large fragments of DNA using yeast artificial chromosomes (YACs) and bacteriophage PI cloning systems.
Key Features
* Spiral bound for easy use at the laboratory bench
*This volume describes:
* Use of methylation-sensitive enzymes for mapping large genomic regions by pulsed field gel electrophoresis
* Step-by-step protocols for the construction, characterisation, screening and use of PI libraries
* Step-by-step protocols for the construction, characterisation, screening and use of YAC libraries
* The analysis of YAC's, their use in chromosome walking, and how to detect and deal with their instability
* Examples on the successful use of YACs for cloning human telomeres, screening cDNA libraries and reconstruction of large genes like that of the Duchenne Muscular Dystrophy gene
* Homologous recombination permitting the insertion of selectable markers into YACs prior to their transfer into mammalian cells
* Automated laboratory procedures for screening libraries and their potential use in generating hybridisation fingertips for genome mapping
* The use of microdissection to generate large number of probes from defined chromosomal regions
* Extensive referencing to recently published primary literature
* Cross referencing between chapters
Key Features
* Spiral bound for easy use at the laboratory bench
*This volume describes:
* Use of methylation-sensitive enzymes for mapping large genomic regions by pulsed field gel electrophoresis
* Step-by-step protocols for the construction, characterisation, screening and use of PI libraries
* Step-by-step protocols for the construction, characterisation, screening and use of YAC libraries
* The analysis of YAC's, their use in chromosome walking, and how to detect and deal with their instability
* Examples on the successful use of YACs for cloning human telomeres, screening cDNA libraries and reconstruction of large genes like that of the Duchenne Muscular Dystrophy gene
* Homologous recombination permitting the insertion of selectable markers into YACs prior to their transfer into mammalian cells
* Automated laboratory procedures for screening libraries and their potential use in generating hybridisation fingertips for genome mapping
* The use of microdissection to generate large number of probes from defined chromosomal regions
* Extensive referencing to recently published primary literature
* Cross referencing between chapters
