Patent Document ID: 8990059
Application ID: 13500431
Patent Flag: 1

Claim One:
1. A computer implemented method for obtaining information for automatic quantification of nucleic acid sequences from an amplification curve of a target nucleic acid sequence obtained from an amplification reaction process, comprising the steps: (a) defining, using a computer, at least one model function that describes the amplification curve and that contains at least one parameter that is related to a physical quantity that has an impact on the signals that are recorded, (b) fitting, using the computer, said model function to the amplification curve, (c) obtaining, using the computer, information with respect to said physical quantity by identifying the value of said at least one parameter that results in a best fit of the model function wherein the at least one parameter is at least one of an absolute initial quantity of the target nucleic acid sequence N0, and parameters E0, α, ε and γ, and wherein the model function is defined as IF(n), IF ⁡ [ n ] = N uq ⁡ [ n ] ⁢ I uq + N q ⁡ [ n ] ⁢ I q 1 + γ · n , wherein 
 N q [n]=N p −N uq [n], and wherein 
 N uq [n]=N[n]+N suq [n], and wherein 
 N suq [n]=N suq [n− 1 ]+β·N q [n− 1], and wherein 
 N[n]=N[n− 1](1 +E[n ]), and wherein E ⁡ [ n ] = E 0 - α ⁡ ( N ⁡ [ n - 1 ] N p - N ⁡ [ n - 1 ] ) ɛ wherein n=cycle number; N(n)=number of target nucleic acid molecules at cycle n; N 0 =N(0) absolute initial number of target nucleic acid sequence; N uq (n)=number of unquenched fluorescent reporters at cycle n; I uq =fluorescence intensity parameter for unquenched fluorescent reporters; N q (n)=number of quenched fluorescent reporters at cycle n; I q =fluorescent intensity parameter for quenched fluorescent reporters; γ=parameter related to the absorption properties of the container used for performing the amplification process; N p =initial number of fluorescent reporter probes; N suq (n)=number of spontaneously unquenched fluorescent reporters at cycle n; β=fraction of fluorescent reporters that unquench spontaneously; E(n)=amplification efficiency of cycle n; E 0 =initial amplification efficiency; α=scaling factor related to inhibition of the amplification process; ε=parameter related to the sensitivity of the amplification process to PCR primer depletion.