Patent ID: 8852867

Claim:
A method for amplifying a polynucleotide sequence complementary to a target polynucleotide sequence, comprising: (a) extending a composite amplification primer in a complex comprising (i) a polynucleotide template comprising the target sequence, wherein the polynucleotide template comprising the target sequence is DNA; and (ii) the composite amplification primer, wherein the composite amplification primer comprises an RNA portion and a 3′ DNA portion, wherein the composite amplification primer is hybridized to the polynucleotide template; (b) cleaving the RNA portion of the annealed composite amplification primer with an enzyme that cleaves RNA from an RNA/DNA hybrid such that another composite amplification primer hybridizes to the template and repeats primer extension and strand displacement, whereby multiple copies of a primer extension product comprising sequence complementary to the target sequence are produced; and (c) incubating the multiple copies of the primer extension product comprising sequence complementary to the target sequence with an enzyme that is capable of cleaving single-stranded RNA, wherein residual single-stranded RNA from the composite amplification primer in the primer extension product comprising sequence complementary to the target sequence is cleaved.