Patent ID: 6248534
Filing Date: 2001-06-19
Classification: C12Q

Abstract:
A method for determining the detection value of a target RNA comprising:(a) preparing at least one target cDNA by reverse transcription of the target RNA and a standard cDNA by reverse transcription of a standard RNA;(b) preparing at least two separate standard samples each containing a different, predefined amount of the standard cDNA and a separate target sample containing an unknown concentration of the target cDNA;(c) adding different adapters, respectively, to each of the samples in excess of that amount necessary to tag all cDNA present in the sample, to obtain adapter-tagged cDNAs;(d) mixing the samples containing the adapter-tagged cDNAs to obtain a mixture;(e) amplifying only the target adapter-tagged cDNA and the adapter-tagged standard cDNA;(f) obtaining separate measurements for each of the amplified cDNAs;(g) determining a relationship between the measurements and relative amounts of amplified standard cDNA from the measurements for the amplified cDNA, the measurements for the amplified and adapter-tagged standard cDNA each being different in accordance with the different predefined amounts of the standard cDNA;(h) applying the measurement for the amplified target cDNA to the determined relationship to obtain a value for the relative amount of the target cDNA; and(i) determining a detection value for the target RNA based on the obtained relative amount value for the target cDNA.