Patent ID: 6858413
Filing Date: 2005-02-22
Classification: C12Q

Abstract:
1. A method of generating multiple copies of a nucleic acid sequence of interest, said method comprising the steps of: (a) hybridizing a composite primer to a target polynucleotide, wherein the composite primer comprises an RNA portion and a 3′ DNA portion, the 3′ DNA portion comprising a 3′ most nucleotide, such that the 3′ most nucleotide of the 3′ DNA portion hybridizes from 1 to about 10 nucleotides from the sequence of interest; (b) extending the composite primer with DNA polymerase under conditions that permit primer extension, whereby a primer extension product is produced; and (c) cleaving the RNA portion of the primer extension product of(b) with an enzyme that cleaves RNA from an RNA/DNA hybrid such that the cleaved primer extension product dissociates from the target polynucleotide, wherein the primer extension product is of a size that when the RNA is cleaved the cleaved primer extension product dissociates from the target polynucleotide under essentially the same conditions as those for primer extension, whereby multiple copies of the sequence of interest are produced.