Application of uMelt SM algorithm in Real-time PCR melting curve analysis

Abstract

One of the challenges in interpreting the results obtained in qPCR is to make sure the amplicons are specific and that the melting curve analysis is used to examine it. Additional peaks in the melting curve is not always indicative of a problem, for this purpose, in this paper, a web-based tool called uMelt SM is suggested to researchers as a practical and simple way for the correct analysis of the melting curve which provides the possibility of predicting the DNA melting curve and denaturation profiles of the high-fluorescence resolution of PCR products. The results of this study showed that the melting curves were generated based on parameters were generated and an appropriate algorithm for working with this software was presented. Finally, actin and superoxide dismutase genes from Pyricularia oryzae were presented as a suitable model for determining the predicted curve using this software and Real-Time PCR curve was also drawn. Results of the uMelt SM predicted melting curves showed a high degree of compliance with the real-time PCR melting curves, which confirms the advantages of ease of use, saving time, cost, and effort in the experimentalist part when using uMelt SM software.

Keywords

• qpcr
• melting curve
• experiment
• predict
• umeltsm