Patent ID: 8198026

Claim:
A method of amplifying specific fragments of a target DNA or RNA with the aid of a polymerase chain reaction in real time, the method comprising: (a) providing a first oligonucleotide primer that comprises a donor fluorescent dye and a second oligonucleotide primer that comprises an acceptor fluorescent dye, wherein the first and second oligonucleotide primers are designed to anneal to partially overlapping portions of a target nucleic acid, wherein such overlap is not more than one nucleotide; (b) allowing the primers to anneal to the target nucleic acid at partially overlapping positions; (c) carrying out a polymerase chain reaction which allows for a fluorescent resonance transfer of energy between the donor dye and the acceptor dye; (d) detecting an increase of fluorescent emission of the acceptor dye; and (e) correlating the increase in the emission of the acceptor dye with the accumulation of the specific fragment of DNA or RNA.