System and method for modeling and subtracting background signals from a melt curve

A method for removing a background signal from a DNA melting curve on a device having at least one DNA sample and which includes a thermal system in communication with the device, the thermal system being controlled to continuously increase a temperature of the at least one DNA sample to cause a DNA melting reaction resulting in denaturing dsDNA to ssDNA, the method comprising:

• (a) generating, by the device performing the DNA melting reaction, a melting curve F(T) for the at least one DNA sample, wherein F is a fluorescence signal indicative of a DNA denaturation process being performed using the thermal system and T is the temperature of the DNA sample;
• (b) providing a mathematical model to fit to the melting curve F(T), the mathematical model represented by a sum of a background reaction term and at least one DNA melting reaction term, each DNA melting reaction term being indicative of the DNA denaturation process being performed by the device and the background reaction term being indicative of the background signal;
• (c) fitting the mathematical model to the melting curve by calculating model parameters for a model curve;
• (d) estimating a maximum difference between the melting curve F (T) and the model curve and a temperature corresponding to the maximum difference;
• (e) iteratively adding one DNA melting reaction term at a time to the sum representing the mathematical model and refitting the sum to the melting curve F (T) to recalculate all model parameters in response to the difference between the melting curve and the model curve estimated in step (d) being greater than a threshold, wherein an initial temperature parameter for each newly added DNA melting reaction term equals to the temperature corresponding to the maximum difference between the melting curve and the model curve obtained in step (d);
• (f) stopping the iteration process of step (e) when the difference between the melting curve and the model curve is less than the threshold;
• (g) subtracting the background reaction term having parameters estimated in the last iteration of step (e) from the melting curve F(T);
• and
• (h) outputting the generated melting curve F(T) based on the subtraction in step (g), wherein the generated melting curve output in step (h) includes information associated with DNA melt reactions without the background reaction terms enabling identification of one or more characteristics associated with the at least one DNA sample undergoing the melting reaction.