Masters Thesis

Fluorescence in situ hybridization to isolated chromosomes in suspension

Fluorescence in situ hybridization (FISH) is a powerful technique that has found a wide range of applications ln modern molecular and cell biology. Rapid advances in FISH have allowed its use in areas ranging from gene mapping and prenatal diagnosis, to characterizations of complex chromosome abnormalities and genome wide DNA alterations associated with various cancers. Because of its many potential applications, this study was designed to develop protocols that will allow for the successful hybridization of fluorescent probes to isolated chromosomes in suspension. Because flow cytometric analysis of the hybridized chromosomes is the next logical step in the development of this technique, work was also done to determine the cell culture and chromosomes isolation procedure which give the optimal flow histogram of the isolated chromosomes and to perform the initial flow cytometric analysis of chromosomes that have been hybridized in suspension.

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