FISH Kits & Probes
Fluorescence in situ hybridization (FISH) is a molecular cytogenetic technique that enables the detection and localization of specific DNA or RNA sequences on chromosomes and nuclei. FISH is a very important molecular technique not only in plant and cell research but also for clinical and medical applications such as tumor diagnosis and prognosis (Oliveira and French 2005), chromosome aberration (Volpi and Bridger 2008) and prenatal chromosomal analysis. The principle of this method is to hybridize small single stranded nuclear acid molecules such as DNA and RNA (probe) to complementary single sequences on chromosomes or nuclei (target), which are immobilized on glass slides. The probes are labeled directly or indirectly using e.g. fluorescent nucleotides. These labeled nucleotides, which carry a covalent bond with a fluorochrome, are synthetically produced. When this fluorophore is excited with light of a specific wavelength meeting the characteristics of the molecule, it emits fluorescence, which can be detected under the microscope. The probes could also be indirectly labeled using molecules, which could be detected through fluorescent antibodies or in a radioactive manner (Volpi and Bridger 2008). After labeling, FISH probes and targets are thermally denatured and hybridized. The standard FISH technique has, however, often some limitations, namely poor signal to background noise ratio due to non-specific binding of probes to the chromosomes or cell surface. In addition, the labeling rate of traditional FISH probes is low. Therefore, routine in situ detection of sequences requires highly labeled and sensitive probes, which are not always available.
The baseclick’s Click-FISH Kits and probes overcome these limitations and provide all reagents required for the generation of highly labeled fluorescence in situ hybridization probes via an easy-to-handle method based on click chemistry.
1. Volpi EV, Bridger JM (2008) FISH glossary: an overview of the fluorescence in situ hybridization technique. Biotechniques 45: 385
2. Oliveira AM, French CA (2005) Applications of fluorescence in situ hybridization in cytopathology - A review. Acta Cytol. 49: 587