Method of processing data in melting of real-time polymerase chain reactions

The derivative melting curve approximating method with derivative SF is suggested. The method is supplemented with zero line drift compensation and the melting curve noise filtration. The method is closer to the traditional methods of value Tm calculating, that are familiar to users, because they usually use melting curve derivative. In constructing derivative of SF zero line instability is completely eliminated. Significant noise reduction is achieved by filtering. The method application results are presented in tabular and graphical forms. The comparison of the proposed methods for the melting point Tm determination and the traditional method is performed. It is recommended to reduce the value of temperature variation step to approximately 0.1 °C when the traditional method is used. The analysis requires considerable time as it is necessary to achieve accurate temperature adjustment at each step. It is shown that step about 1 °C can be used the proposed method is used, thus saving considerable analysis time and the melting point can be calculated with a resolution not worse than 0.1 °C.