In situ hybridization (ISH) is used to visualize defined nucleic acid sequences in cellular preparations by hybridization of complementary probe sequences. Probe sequences can be labeled with isotopes, but nonisotopic ISH is used increasingly as it is considerably faster, usually has greater signal resolution, and provides many options to simultaneously visualize different targets by combining various detection methods. The most popular protocols use fluorescence detection, as described here. These protocols have many applications, from basic gene mapping and diagnosis of chromosomal aberrations1,2,3 to detailed studies of cellular structure and function, such as the painting of chromosomes in three-dimensionally preserved nuclei4,5. This protocol describes fluorescence in situ hybridization (FISH) of biotin- or digoxigenin-labeled probes to denatured metaphase chromosomes and interphase nuclei. The hybridized probes are detected and visualized using fluorochrome-conjugated reagents.
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References
Lichter, P. & Ward, D.C. Is non-isotopic in situ hybridization finally coming of age? Nature 345, 93–95 (1990).
Lichter, P., Boyle, A.L., Cremer, T. & Ward, D.C. Analysis of genes and chromosomes by nonisotopic in situ hybridization. Genet. Anal. Tech. Appl. 8, 24–35 (1991).
Bentz, M. et al. Detection of trisomy 8 on blood smears using fluorescence in situ hybridization. Leukemia 7, 752–757 (1993).
Zirbel, R.M., Mathieu, U.R., Kurz, A., Cremer, T. & Lichter P. Evidence for a nuclear compartment of transcription and splicing located at chromosome domain boundaries. Chromosome Res. 1, 93–106 (1993).
Kurz, A. et al. Active and inactive genes localize preferentially in the periphery of chromosome territories. J. Cell Biol. 135, 1195–1205 (1996).
Lichter, P., Cremer, T., Borden, J., Manuelidis, L. & Ward, D.C. Delineation of individual human chromosomes in metaphase and interphase cells by in situ suppression hybridization using recombinant DNA libraries. Hum. Genet. 80, 224–234 (1988).
Pinkel, D. et al. Fluorescence in situ hybridization with human chromosome-specific libraries: Detection of trisomy 21 and translocations of chromosome 4. Proc. Natl. Acad. Sci. USA 85, 9138–9142 (1988).
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Fluorescence in situ hybridization. Nat Methods 2, 237–238 (2005). https://doi.org/10.1038/nmeth0305-237
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DOI: https://doi.org/10.1038/nmeth0305-237
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