An embodiment of the invention discloses new methods for designing labeled
nucleic acid probes and primers by labeling oligonucleotides with a
plurality of spectrally identical or similar dyes and optionally with one
or more quencher dyes. Oligonucleotides labeled in accordance with some
embodiments of the invention exhibit a detectable increase in signal, for
example, fluorescent signal when the labeling dyes are separated from one
another. Methods for separating the dye include cleaving the labeled
oligonucleotides include using enzymes that have 5'-exonuclease activity.
In one embodiment nucleic acid primers of the present invention may
fluoresce upon hybridization to a target sequence and incorporation into
the amplification product. Nucleic acid probes and primers of the present
invention have wide applications ranging from general detection of a
target nucleic acid sequence to clinical diagnostics. Major advantages of
the oligonucleotides including nucleic acid probes and primers of many
embodiments of the present invention are their synthetic simplicity,
spectral versatility and superior fluorescent signal.