Chromosome Analysis Protocols

ISBN-13: 9780896032439 / Angielski / Miękka / 1994 / 509 str.

John R. Gosden

Opis

Chromosomes, as the genetic vehicles, provide the basic material for a large proportion of genetic investigations, from the construction of gene maps and models of chromosome organization, to the inves tigation of gene function and dysfunction. The study of chromosomes has developed in parallel with other aspects of molecular genetics, beginning with the first preparations of chromosomes from animal cells, through the development of banding techniques, which permitted the unequivocal identification of each chromosome in a karyotype, to the present analytical methods of molecular cytogenetics. Although some of these techniques have been in use for many years, and can be learned relatively easily, most published scientific reports as a result of pressure on space from editors, and the response to that pressure by authors contain little in the way of technical detail, and thus are rarely adequate for a researcher hoping to find all the necessary information to embark on a method from scratch. A new user needs not only a detailed description of the methods, but also some help with problem solving and sorting out the difficulties en countered in handling any biological system. This was the require ment to which the series Methods in Molecular Biology is addressed, and Chromosome Analysis Protocols forms a part of this series."

Recenzje

...a very worthwhile book...invaluable for those trying to venture into the area of molecular cytogenetics...Provides a ready source of both old and new cytogenetic methods and contains a wealth of troubleshooting tips from experienced workers in the field.-Modern Pathology

Spis treści

Mitotic Metaphase Chromosome Preparation from Peripheral Blood for High Resolution. Chromosome Preparation from Hematological Malignancies. Meiotic Chromosome Preparation. Preparation of Chromosomes for Scanning Electron Microscopy. Immortalized Cell Lines: Chromosome Preparation and Banding. Chromosome Banding and Identification: Absorption Staining. Chromosome Banding and Identification: Fluorescence. Chromosome Banding: Stain Combinations for Specific Regions. Inhibition of Chromosome Condensation. Analysis of Chromosomes with Restriction Endonucleases and DNase Hypersensitivity. Automated Karyotype Analysis. Bivariate Chromosome Analysis Using a Commercial Flow Cytometer. Chromosome Sorting by Flow Cytometry: Production of DNA Libraries and Gene Mapping. Electrophoretic Karyotype Analysis: Pulse Field Gel Electrophoresis. Immunofluorescence Techniques Applied to Mitotic Chromosome Preparations. Immunocytochemical Techniques Applied to Meiotic Chromosome Preparations. Immortalized Cell Lines: Their Creation and Use in Gene Mapping. Oligonucleotide-Primed In Situ DNA Synthesis: An Alternative to In Situ Hybridization for Gene Mapping and the Investigation of Genome Organization. Electron Microscopic Localization of In Situ Hybrids. Protocols for Chromosome Mediated Gene Transfer: Selection Strategies, Transgenome Analysis, Enrichment Cloning, and Mapping. Construction of Chromosome Specific Libraries of Yeast Artificial Chromosome Recombinants from Somatic Hybrid Cell Lines. Yeast Artificial Chromosome Recombinants in a Global Strategy for Chromosome Mapping: Amplification of Internal and Terminal Fragments by PCR, and Generation of Fingerprints. Chromosome Dissection and Cloning. Molecular Analysis of Chromosome Aberrations in Hematological Malignancies: Southern Hybridization. Molecular Analysis of Chromosome Aberrations: In Situ Hybridization. Alu- and L1-Primed PCR-Generated Probes for Non-Isotopic In Situ Hybridization. Chromosome Substructure Investigation:Telomeres. Index.