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Molecular Biology > In-situ Hybridization
The use of a DNA or RNA probe to detect the complementary sequence.
In situ hybridization is like all nucleic acid hybridization in being a technique in which single-stranded nucleic acids (DNA or RNA) are permitted to interact so that complexes, or hybrids, are formed by molecules with sufficiently similar, complementary sequences. Through nucleic acid hybridization, the degree of sequence identity can be determined and specific sequences detected and sometimes those sequences can be located on a specific chromosome.
In situ hybridization is done for this purpose. It involves hybridizing a labelled nucleic acid (often labelled with a fluorescent dye) to suitably prepared cells or histological sections. This is used particularly to look for specific transcription or localization of genes to specific chromosomes (via fluorescent in situ hybridization, or FISH analysis).
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Fluorescence In Situ Hybridization (FISH)
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Fluorescence in situ hybridization (FISH) uses fluorescent molecules to vividly paint genes or chromosomes.
This technique is particularly useful for gene mapping and for identifying chromosomal abnormalities.
FISH involves the preparation of short sequences of single-stranded DNA, called probes, which are complementary to the DNA sequences the researchers wish to paint and examine.
These probes hybridize, or bind, to the complementary DNA and, because they are labeled with fluorescent tags, allow researchers to see the location of those sequences of DNA.
Unlike most other techniques used to study chromosomes, which require that the cells be actively dividing, FISH can also be performed on nondividing cells, making it a highly versatile procedure.
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