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        Fluorescence In Situ Hybridization (FISH) to Metaphase and Interphase Chromosomes

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        The unambiguous identification of human chromosomes became possible with the discovery and implementation of G-banding techniques (1 ). Almost immediately, investigators developed various methods to physically map specific DNA sequences to banded chromosomes. A commonly used early technique involved the hybridization in situ of radioactively labeled probes to heat-denatured human metaphase chromosomes (reviewed in 2 ). These techniques were efficient, yet costly, time-consuming, and technically difficult. Isotopic hybridization in situ was rapidly superseded by nonisotopic techniques—especially those utilizing fluorescently labeled probes (3 6 ). This chapter describes basic methodology for the accomplishment of metaphase and interphase fluorescence in situ hybridization (FISH).
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