Principles of fluorescence in situ hybridization | 合法醫療器材資訊網

Principlesoffluorescenceinsituhybridization.(a)ThebasicelementsofFISHareaDNAprobeandatargetsequence.(b)Beforehybridization,theDNA ...

Principles of fluorescence in situ hybridization

(a) The basic elements of FISH are a DNA probe and a target sequence. (b) Before hybridization, the DNA probe is labeled by various means, such as nick translation, random primed labeling, and PCR. Two labeling strategies are commonly used: indirect labeling (left panel) and direct labeling (right panel). For indirect labeling, probes are labeled with modified nucleotides that contain a hapten, whereas direct labeling uses nucleotides that have been directly modified to contain a fluorophore. (c) The labeled probe and the target DNA are denatured. (d) Combining the denatured probe and target allows the annealing of complementary DNA sequences. (e) If the probe has been labeled indirectly, an extra step is required for visualization of the nonfluorescent hapten that uses an enzymatic or immunological d...