Nucleic acids analyzer holes temperature spread determining method

The temperature determining methods application results based on fluorescence signals approximation for the ANK-96 analyzer are considere d. An improved method for determining the temperature dispersion in the wells is proposed on the base of the fluorescence signals nonlinear approximation by sigmoidal function (SF). Fluorescent dyes with a high temperature dependence (about 15 % K-1) are used. The results are presented in tabular and graphical form. It was shown that the melting temperature of the DNA of all the tubes and their average value correspond to the absolute temperature scale in °C. The deviation of temperatures is not more than ± 0.2 °С. The average value of the melting points of the DNA in all tubes is T mc = 80.82 °C. The maximum temperature deviation in 6 groups of 16 tubes (4 × 4) from the average value is 0.15 °C and does not exceed the maximum allowable values ± 0.2 °C. It is suggested to use the method based on nonlinear approximation by sigmoid function (SF) in tuning, calibrating and verifying the thermal parameters of nucleic acid analyzers.