Detection of DNA hybridization using induced fluorescence resonance energy transfer.
Induced fluorescence resonance energy transfer (iFRET) is a variation of resonance energy transfer that is particularly well-suited for the detection of DNA hybridization. The underlying mechanism involves monitoring changes in fluorescence that are the result of an energy transfer reaction between a specific pair of donor and acceptor moieties. In iFRET, the donor is a dye that only fluoresces while interacting with double-stranded DNA and the acceptor is dye that is covalently linked to an oligonucleotide probe. Hybridization of the probe to its complement induces excitement of the donor dye and subsequent energy transfer to the acceptor dye. The energy transfer reaction (and concomitant hybridization status) can easily be followed by monitoring the fluorescence output of the acceptor dye. Because the interaction of the donor dye is reversible and dependent on the presence of double-stranded DNA, iFRET is extremely useful and herein demonstrated in the generation of DNA melting curves.
Howell
W Mathias
WM
Center for Genomics and Bioinformatics, Karolinska Institute, Stockholm, Sweden.
D004247:DNA; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D005456:Fluorescent Dyes; D015336:Molecular Probe Techniques; D015335:Molecular Probes; D009693:Nucleic Acid Hybridization / Q000379:methods*
D005456:Fluorescent Dyes; D015335:Molecular Probes; D004247:DNA