Patent ID: 11879153
Assignee: BIOVUE TECHNOLOGY LTD
Field: Biotechnology (Chemistry)
Classification: CPC C | IPC C

Claim 9:
10. The method of claim 1, said PCR reaction system further comprising:
(vi) a second primer pair for amplifying a second target sequence to generate a second amplicon, said second primer pair comprising a second blocked primer which comprises a second 2′,3′-dideoxyribonucleotide located at the 3′ end of the second blocked primer, wherein said second 2′,3′-dideoxyribonucleotide blocks the extension of said nucleic acid polymerase or combination of nucleic acid polymerases when said second 2′,3′-dideoxyribonucleotide does not anneal to said second target sequence or said second amplicon, and
(vii) a second fluorescent probe complementary to the second target sequence or the second amplicon, said second fluorescent probe comprising a third nucleotide linked to a second fluorophore and a fourth nucleotide linked to a second quencher, wherein the fluorescence signal of said second fluorophore is quenched by said second quencher when said third nucleotide is not hydrolyzed from said probe, and wherein said nucleic acid polymerase or combination of nucleic acid polymerases is capable of hydrolyzing said third nucleotide from said second fluorescent probe bound to said target sequence or amplicon during extension such that the fluorescence signal of said second fluorophore is not quenched by said second quencher.