ASYMMETRIC PCR WITH NUCLEASE-FREE POLYMERASE OR NUCLEASE-RESISTANT MOLECULAR BEACONS

摘要

The invention provides methods for performing combined asymmetric amplification (e.g. asymmetric PCR amplification) and detection of nucleic acid targets using molecular beacons to detect the products. Methods using a polymerase having reduced or eliminated 5' to 3' nuclease activity are provided, as are methods using nuclease-resistant molecular beacons. Asymmetric amplifications using nuclease-free polymerase or nucleease-resistant molecular beacons provide dramatic improvements in signal intensity detected as a result of molecular beacon binding to a target nucleic acid, e.g., during asymmetric PCR. Attendant compositions, systems, devices and kits are also features of the invention.