Patent ID: 6358679
Filing Date: 2002-03-19
Classification: C12Q,Y10S

Abstract:
A method for detecting a target polynucleotide sequence comprising the steps of:(a) amplifying a target polynucleotide with primer extension reagents in a first set of one or more vessels and amplifying an external control polynucleotide with the primer extension reagents in a second set of one or more vessels; wherein the primer extension reagents include a forward primer, a reverse primer, one or more detectable probes, a polymerase, and one or more nucleotide 5â€²-triphosphates; the forward primer and the detectable probe are separated by 0 to 5 nucleotides when hybridized to the external control polynucleotide, or its complement, and the reverse primer and the detectable probe are separated by 0 to 5 nucleotides when hybridized to the external control polynucleotide, or its complement; and the external control polynucleotide begins the amplification process as a single-stranded polynucleotide; and the external control polynucleotide is shorter than the target polynucleotide; and (b) detecting a signal from said one or more detectable probes.