Patent ID: 6551783
Filing Date: 2003-04-22
Classification: C12Q

Abstract:
A method of quantitation of expression of a first and a second target protein-encoding nucleic acids using a PCR based assay in which amplification of nucleic acid is detected by release of a reporter fluorescent signal which is measured as a normalised reporter value (â€œ&Dgr;Rnâ€ ), the method comprising:providing a limiting concentration of a first primer pair, the first primer pair being adapted for amplification of a desired first target, the limiting concentration being selected to provide a first &Dgr;Rn value as measured by release of a first fluorescent reporter compound from a first reporter probe; selecting a non-limiting concentration of a second primer pair and probe set, the second primer pair being adapted for amplification of a desired second target, the amplification being measured by release of a second fluorescent reporter compound from a second reporter probe, the concentration being selected to provide a second &Dgr;Rn value of from three to forty times that provided by said first reporter compound; bringing together said first and second primer pairs and probes at their limited and selected concentrations respectively into contact with a sample of nucleic acid; performing a polymerase chain reaction under conditions in which the release of the first and second reporter compounds is measured; and quantitating the amount of first and second target nucleic acids.