Patent ID: 8163484

Claim:
A method for determining the melting temperature (T m ) of an intermolecular hybridization complex, the method comprising: (a) providing: (i) a probe comprising a light emitting moiety; (ii) a hybridization target that is complementary or partially complementary to said probe wherein said probe and said hybridization target are two separate nucleic acid molecules; (iii) a soluble light emission modifier comprising a thiazine dye or a diazine dye, wherein said soluble light emission modifier is capable of quenching said light emitting moiety; (b) annealing said probe with said hybridization target under conditions wherein base-pairing occurs to form an intermolecular target hybridization complex; (c) altering the temperature of said intermolecular target hybridization complex in the presence of said soluble light emission modifier and measuring an emission of said light emitting moiety; (d) correlating said measured emission of said light emitting moiety with the presence of said intermolecular target hybridization complex as a function of temperature, thereby determining Tm of said intermolecular target hybridization complex based on said measured emission.