Generi Biotech - Compounds and Services for Molecular Biology

• Real-Time PCR Probes
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About Mono Labelled Probes

Functional principle of mono labelled probes (known also as FRET probes) is based on the phenomenon called FRET (Fluorescence Resonance Energy Transfer). It is a photophysical phenomenon with energy transfer occurring between two fluorophores which are in close proximity. If the emission spectrum of one fluorophore (a so called donor) overlaps with the absorption spectrum of the other fluorophore (acceptor), fluorescence intensity of the donor molecule decreases and irradiated energy is captured by the acceptor molecule in the form of fluorescent radiation with longer wavelength.

The system of mono labelled probes consists of two oligonucleotides where one is marked with the donor fluorophore at the end 3´ and the other with an acceptor fluorophore at the end 5´. The probes are designed in such a way so that they hybridize adjacent to each other on the DNA template, formed by the PCR product. If the probes hybridize with a target sequence, energy transfer between fluorophores of the donor-acceptor pair occurs, and fluorescent activity of the acceptor fluorophore increases, which is subsequently measured.

FRET probes provide the advantage of the possibility to perform (in connection with melting curve analysis) detection of SNP polymorphisms and mutation analysis in general.

GENERI BIOTECH supplies mono labelled probes in combination with FAM/Cy5.