Experimental determination of polymerase chain reaction amplification parameters of nucleic acid analizer

The purpose of this article is to identify the conditions that allow to achieve the required test results in minimum time. The method of determination of the real-time polymerase chain reaction (RT-PCR) amplification basic parameters (PCR efficiency and the thermal cycle duration) was suggested. An example of the threshold cycles measuring and calculation of the PCR efficiency of test samples with fluorescent dyes FAM and ROX using breeding techniques. The PCR efficiency measuring error and PCR efficiency dependence on the temperature cycle duration were determined. As a result of the experiments and calculations, the temperature mode with the maximum PCR efficiency closed to the most achievable value was determined. The most effective temperature conditions selection and the cycle time reducing can significantly cut down on the performance of the analysis (it is decreased by 19 % in the experiment).