| CPC C12Q 1/686 (2013.01) | 26 Claims |
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1. A method for performing polymerase chain reaction (PCR), comprising
thermocycling a PCR mixture having a first volume in one amplification zone of an amplification container for a first number of cycles, each of the first number of cycles comprising a first cycle time, a first annealing temperature, a first annealing hold, a first denaturation temperature, and a first denaturation hold,
reducing the volume of the PCR mixture in the same one amplification zone to a second volume, the second volume being smaller than the first volume, and
thermocycling the second volume of PCR mixture in at least a portion of the one amplification zone for a second number of cycles, each of the second number of cycles comprising a second cycle time, a second annealing temperature, a second annealing hold, a second denaturation temperature, and a second denaturation hold, the second cycle time being shorter than the first cycle time and the second denaturation temperature being lower than the first denaturation temperature after reducing the volume of the PCR mixture;
further comprising performing an initial cycle in the one amplification zone prior to thermocycling for a first number of cycles, the initial cycle comprising an initial cycle time, an initial annealing temperature, an initial denaturation temperature, and an initial denaturation hold, wherein the initial denaturation hold is longer than the first denaturation hold, and the initial denaturation temperature is lower than the first denaturation temperature.
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