Patent ID: 6691041
Filing Date: 2004-02-10
Classification: C12Q,G16Z

Abstract:
A method for quantifying a target nucleic acid relative to a reference nucleic acid comprising the steps of:a) determining the amplification efficiencies of the target nucleic acid and of the reference nucleic acid under defined amplification conditions; b) amplifying the target nucleic acid contained in a sample and the reference nucleic acid contained in said sample under said defined amplification conditions; c) measuring the amplification of the target nucleic acid and of the reference nucleic acid in real time; d) setting a defined signal threshold value; e) determining the cycle numbers at which the signal threshold value is exceeded in each case during the amplification of the target nucleic acid and the reference nucleic acid; and f) calculating the original ratio of target nucleic acid to reference nucleic acid according to the following formula N(T)0/N(R)0=E(R)n(R)/E(T)n(T), in whichN(T)0=the original amount of target DNA N(R)0=the original amount of reference DNA E(R)=the amplification efficiency of the reference nucleic acid n(R)=the cycle number of the reference nucleic acid measured in step e) E(T)=the amplification efficiency of the target nucleic acid n(T)=the cycle number of the target nucleic acid measured in step e).