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In situ hybridization
Last reviewed: 04.07.2025
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The hybridization procedure can be carried out not only on a gel, on filters or in solution, but also on histological or chromosomal preparations. This method is called in situ hybridization. A variant of the method, in which DNA or RNA preparations labeled with fluorochromes are used as probes, is called FISH (fluorescein in situ hybradization). The labeled DNA probe is applied to preparations of differentially stained and prepared for hybridization (denatured) metaphase chromosomes. Preliminary treatment of the chromosomes is used to facilitate access of the probe to the genomic DNA. After washing away unbound DNA molecules and applying a light-sensitive emulsion (when using a radioactive label), or performing appropriate treatment (when using biotin- or fluorescein-labeled DNA probes), the localization sites of DNA sequences complementary to the DNA probe used can be directly detected by microscopy as characteristic dots above the corresponding sections of certain chromosomes.
This research method allows us to determine not only the chromosomal affiliation, but also the intrachromosomal localization of the gene being studied.
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