Patent ID: 6248518
Filing Date: 2001-06-19
Classification: C12Q

Abstract:
A method for detecting point mutations in a selected target DNA sequence, wherein the sequence has a reference melting point at which it becomes detached from a complementary sequence, the method comprising:providing a probe having a sequence complementary to the target DNA sequence;attaching at least two dyes to the probe, the dyes producing FRET emissions, wherein the FRET emissions change in response to a change in proximity of the dyes to each other,allowing the probe to hybridize with the DNA sequence to form a probe/target DNA duplex;recording an attached value of the FRET emissions from the dyes of the probe/DNA duplex;heating the probe/DNA duplex;monitoring the temperature at which the FRET emissions from the probe and duplex are reduced to a detached value as a result of the change in the proximity of the dyes to each other; anddetermining an observed melting point of the probe/target DNA duplex by recording the temperature of the probe/DNA duplex at the midpoint between the attached value of the FRET emissions and the detached value of the FRET emissions;whereby if the observed melting point is lower than the reference melting point at least one point mutation exists.