Patent ID: 8852863

Claim:
Method for amplifying and detecting nucleic acid sequences in a reaction cartridge comprising the following steps: (i) providing a sample comprising at least one nucleic acid molecule and a throw-away microfluidic cartridge, (ii) in a first reaction chamber of the cartridge providing reagents for an amplification reaction, (iii) mixing the sample with the amplification reagents, (iv) amplifying the at least one nucleic acid in the first reaction chamber of the cartridge, (v) transferring at least parts of the amplification reaction into a second and third reaction chamber of the cartridge each comprising a probe set, wherein a) each probe set consists of at least three probes, b) each of the probes is specific for a nucleic acid sequence; c) there are at least two probes in each set which carry an identical label, wherein the label is a fluorescent label, d) each of the probes in a given probe set that carries an identical label has a melting temperature (T m ) which differs by more than 2° C. from the other probe in said probe set with the same label, e) wherein the probes carrying the identical label differ in melting temperature (T m ) in a way that they are distinguishable by melting point, (vi) performing a melting point analysis in the second and/or third reaction chamber in order to determine which of the probes has specifically bound a nucleic acid, wherein the amplification and melting point analysis takes place when the cartridge is located in an amplification device, and wherein the cartridge comprises a connection between the first reaction chamber and the second and/or third reaction chamber.