The present invention is directed to a method for performing a one-step RT-PCR for amplifying a target RNA comprising the steps (i) providing a sample which is supposed to contain said target RNA (ii) adding a reaction mixture comprising all reagents necessary to reverse transcribe said target RNA into cDNA and amplify at least a portion of said cDNA (iii) incubating said sample for a time interval of 0 seconds to 40 seconds at a temperature between 20° C. and 65° C., and (iv) subjecting said sample to multiple cycles of a thermocycling protocol wherein the temperature of said sample is varied between at least a first temperature between 37° C. and 72° C. and a second temperature between 85° C. and 100° C.
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