System and method for melting curve normalization

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 that continuously increases 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 a melting curve F(T) for a DNA sample, wherein F is a fluorescence signal indicative of a DNA denaturation process and T is the temperature of the DNA sample;
• (b) dividing the melting curve into three regions, the three regions being a pre-DNA melt region [T1, T2] and a post-DNA melt region [T3, T4] corresponding to a background reaction, and a central melt region [T2, T3] corresponding to the DNA melting reaction;
• (c) fitting a background reaction model to the regions [T1, T2] and [T3, T4] of the melting curve by determining parameters of the background reaction model and calculating a background reaction curve;
• and
• (d) subtracting the background reaction curve from the melting curve F(T) at the temperature interval [T1, T4].

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 that continuously increases 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 a melting curve F(T) for a DNA sample, wherein F is a fluorescence signal indicative of a DNA denaturation process and T is the temperature of the DNA sample;
• (b) transforming the melting curve F(T) to a curve log(F(T)−u) for different values of a u factor;
• (c) for each value of the u factor, fitting a first background model B1(T)=s1T+t1 to a pre-DNA melt region [T1, T2] of the transformed melting curve log(F(T)−u) and a second background model B2(T)=s2T+t2 to a post-DNA melt region [T3, T4] of the transformed melting curve log(F(T)−u) thereby determining s1, s2, t1, t2, T1, T2, T3, and T4;
• (d) selecting the factor for which s1=s2;
• and
• (e) removing the background signal from the melting curve F(T) based upon the first and second background models B1(T) and B2(T).

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 that continuously increases 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 a melting curve F(T) for a DNA sample, wherein F is a fluorescence signal indicative of a DNA denaturation process and T is the temperature of the DNA sample;
• (b) transforming the melting curve F(T) to a curve log(F(T)−u) for a first approximation of a u factor;
• (c) calculating a first background model B1(T)=sT+t1 for a predefined pre-DNA melt region [T1, T2] of the transformed melting curve log(F(T)−u) and a second background model B2(T)=st+t2 for a predefined post-DNA melt region [T3, T4] of the transformed melting curve log(F(T)−u) by minimizing a first objective function to calculate parameters s, t1, and t2;
• (d) taking s, t1, and t2 calculated in step (c) as an approximation to calculate a second approximation of the u factor, the second approximation for the u factor being calculated by minimizing a second objective function;
• (e) repeating steps (c) and (d) until the parameters u, t1, and t2, and s converge, wherein u, t1, and t2, and s ultimately minimize the first and second objective functions;
• and
• (f) removing the background signal from the curve F(T) based upon the first and second background models B1(T) and B2(T).

A method comprising:

• ramping a temperature of a plurality of DNA samples in a DNA vessel by using a temperature control system to achieve DNA denaturing, each of the plurality of DNA samples having an internal template control having a pre-determined melt temperature;
• generating a plurality of DNA melting curves for the plurality of samples by using an imaging system to measure fluorescence as a function of temperature and calculating a negative derivative plot for each generated DNA melting curve;
• and
• calculating, by a controller, start and end temperature values, ITC1 and ITC2, defining an internal template control (ITC) region in the derivative plot of the plurality of DNA melting curves.

A method for normalizing a plurality of DNA melting curves by determining a melt region in the plurality of DNA melting curve, the method comprising:

• ramping a temperature of a plurality of DNA samples in a DNA vessel by using a temperature control system to achieve DNA denaturing, each of the plurality of DNA samples having an internal template control having a pre-determined melt temperature;
• generating a plurality of DNA melting curves for the plurality of samples by using an imaging system to measure fluorescence as a function of temperature;
• and
• identifying, by a controller, start and end temperatures of the melt region in the plurality of melting curves;
• and
• normalizing, by the controller, the plurality of DNA melting curves based at least in part on the identified melt region of the melting curves.

A method comprising:

• receiving, at a server, raw experimental data from a client machine, the experimental data having a format defining an instrument where the raw experimental data was generated;
• and
• processing, at the server, the received raw experimental data to generate processed experimental;
• obtain, by the client computing device, the processed experimental data;
• and generating a user interface on the client machine that reflects a type of the instrument that generated the raw experimental data.

A server comprising:

• one or more processors;
• a storage including instructions that, when executed by the one or more processors, configures the server to:
• receive raw experimental data including format information identifying an instrument that was used to generate the raw experimental data;
• store the raw experimental data in the storage;
• generate processed experimental data using the stored raw experimental data including perform one or more processing operations on the raw experimental data using the format information to generate a data object usable to generate a user interface that provides a visual depiction of a sample plate used in the instrument that generated the raw experimental data;
• and
• transmit the data object representing the processed experimental data to a client computing device for outputting the user interface generated based on the transmitted data object.

A method comprising:

• receiving, at a server, raw experimental data including format information identifying an instrument that was used to generate the raw experimental data;
• storing the raw experimental data in a storage of the server;
• generating, at the server, processed experimental data using the stored raw experimental data including perform one or more processing operations on the raw experimental data using the format information to generate a data object usable to generate a user interface that provides a visual depiction of a sample plate used in the instrument that generated the raw experimental data;
• and
• transmitting the data object representing the processed experimental data to a client computing device for outputting the user interface generated based on the transmitted data object.

A computing device comprising

• one or more processors;
• a storage including instructions that, when executed by the one or more processors, configures the server to:
• provide, to a server, raw experimental data including format information identifying an instrument that was used to generate the raw experimental data;
• receive, from the server, a data object representing processed experimental data that was generated using the stored raw experimental data and which includes processed format information;
• generating a user interface using the processed format information that provides a visual depiction of a sample plate used in the instrument that generated the raw experimental data;
• and
• displaying the generated user interface on a display screen.

A computing device comprising

• one or more processors;
• a storage including instructions that, when executed by the one or more processors, configures the server to:
• provide, to a server, raw experimental data including format information identifying an instrument that was used to generate the raw experimental data;
• receive, from the server, a data object representing processed experimental data that was generated using the stored raw experimental data and which includes processed format information;
• generating a user interface using the processed format information that provides a visual depiction of a sample plate used in the instrument that generated the raw experimental data;
• and
• displaying the generated user interface on a display screen.

A method comprising

• providing, by a client computing device to a server, raw experimental data including format information identifying an instrument that was used to generate the raw experimental data;
• receive, at the client computing device from the server, a data object representing processed experimental data that was generated using the stored raw experimental data and which includes processed format information;
• generating, by the client computing device, a user interface using the processed format information that provides a visual depiction of a sample plate used in the instrument that generated the raw experimental data;
• and
• displaying, at the client computing device, the generated user interface on a display screen.