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Probes for Molecular Cytogenetics According to Their Labeling
- Jan 30, 2019 -

When FISH era started, there was only one possibility to label the applied probes, i.e., incorporate a biotin-labeled nucleotide analogue into the probe DNA and detect it after hybridization by avidin or streptavidin coupled to a fluorescence molecule—at that time most often FITC was applied in one-color FISH approaches . This labeling method was called “indirect labeling,” as the fluorochrome was not directly attached to the probe DNA. Yet, routinely applied as “indirectly labeled probes” are only biotin and digoxigenin, which can be detected by antibodies specifically tagged with different fluorochromes; besides also dinitrophenol may be used besides biotin or digoxigenin as hapten. Nowadays there are dozens of fluorochromes available on the market which can be purchased as already attached to a nucleotide analogue as well as detected by suited filters .

Reference : Fluorescence In Situ Hybridization (FISH) Thomas Liehr 


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