Patent ID: 7226732
Filing Date: 2007-06-05
Classification: C12Q,G16B,Y10T

Abstract:
1. A method of determining if a probe is sufficiently intact to detect a target nucleic acid in a nucleic acid amplification reaction, the method comprising: (a) providing a mixture comprising at least one probe, which probe is capable of hybridizing to the target nucleic acid; (b) measuring a signal of the probe at one or more time points when the probe is not bound to the target nucleic acid; and (c) determining the integrity of the probe by comparing the signal of the probe at the one or more time points with at least one pre-determined threshold value indicative of the signal of the probe when the probe is intact, wherein: the signal is fluorescence, the probe is capable of hybridizing with a target nucleic acid, the probe comprising a fluorophore and a quenching agent, wherein heating the probe causes the probe to change conformation from a first self-annealing conformation to a second conformation, thereby changing the distance between the fluorophore and the quenching agent such that the fluorescence of the fluorophore is quenched or altered in the first conformation compared to the fluorescence of the fluorophore in the second conformation, and the fluorescence of the probe is measured at a temperature equal to or below the self-annealing temperature of the probe.