| abstract |
Disclosed herein are methods of detecting target nucleic acids. In particular, methods for avoiding loss of the fluorescent label form an amplicon that is generated using a Scorpion primer and a polymerase with 5' exonuclease activity. The methods use a Scorpion primer which comprises a fluorophore, a quencher, and in 5? to 3? order, a probe region, a linker region and a primer region, wherein the quencher is located at or near the 5? end, and, wherein the primer is complementary to the target nucleic acid and the probe region hybridizes to a complementary sequence in an extension product of the primer. The methods provide for detection of target nucleic acids in simplex or multiplex formats. |