Abstract
Fluorescence in situ hybridization (FISH) can be used to localize specific DNA sequences on metaphase chromosomes, interphase nuclei, and experimentally extended DNA or chromatin fibers. Depending on the hybridization target, FISH techniques show widely different levels of DNA resolution. Mechanically stretched or elongated chromosomes fill the resolution gap between metaphase FISH and fiber FISH, allowing the rapid and straightforward ordering and localization of clones along the length of an entire chromosome with a 100- to 200-kb resolution. Although various genome projects have provided very high-resolution physical maps of human and important animal genomes, FISH is still an exceptionally versatile mapping tool. The combined application of protein staining by immuno-fluorescent techniques and DNA staining by FISH provides a powerful tool for mapping chromosomal proteins to specific DNA sequences. The most suitable sources of mitotic cells are long-term in vitro cell cultures such as EBV-transformed lymphoblasts, fibroblasts, or established cell lines.
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