abstract |
A method for identifying a plurality of target nucleic acid molecules in a sample. The method provides a plurality of oligonucleotide probe sets. Each set comprises a first and a second probe, each having a target-specific portion and a tunable portion with an acceptor or a donor group. The first probe further comprises an endcapped hairpin. A reaction comprises a denaturation and hybridization cycle. Under the hybridization, the set of probes hybridize in a base-specific manner to their respective target nucleotide sequences, and ligate to one another to form a ligation product. Under conditions that permit hybridization of the tunable portions of the ligation product to one another, an internally hybridized ligation product formed, which allows the detection of the fluorescence resonance energy transfer (FRET). A method comprising PCR amplification is also disclosed. |