Patent ID: 8183359
Filing Date: 2012-05-22
Classification: C12P,C12Q

Abstract:
1. A kit for use in amplifying a DNA target sequence, said kit comprising: a priming oligonucleotide which hybridizes to the 3′-end of a DNA target sequence such that a primer extension reaction can be initiated therefrom; a promoter oligonucleotide comprising first and second regions, wherein said first region has a base sequence which corresponds to a region at the 5′-end of said target sequence and which hybridizes to a 3′-region of a DNA primer extension product comprising said priming oligonucleotide, wherein said second region is a promoter for an RNA polymerase and is situated 5′ to said first region, and wherein said promoter oligonucleotide is modified to prevent the initiation of DNA synthesis therefrom; and a displacer oligonucleotide which hybridizes to a target nucleic acid containing said target sequence upstream from said priming oligonucleotide such that a primer extension reaction can be initiated therefrom, provided that any oligonucleotide included in said kit that comprises a promoter for an RNA polymerase is modified to prevent the initiation of DNA synthesis therefrom, provided that said kit does not include a restriction endonuclease, and further provided that said priming oligonucleotide does not include an RNA region which hybridizes to said target nucleic acid and which is selectively degraded by an enzyme activity when hybridized to said target nucleic acid.