Patent ID: 7344834

Claim:
A method for amplifying a target nucleic acid sequence comprising the steps of; a) forming a nucleotide amplification reaction mixture comprising a DNA template containing a target nucleic acid sequence; a single chimeric oligonucleotide primer consisting of a deoxyribonucleotide sequence with a ribonucleotide base at the 3′ terminus that binds to said DNA template; a non-extendable oligonucleotide blocker that binds to said DNA template downstream from said primer; a DNA polymerase which lacks 5′ exonuclease activity; and a double-strand-specific ribonuclease, and appropriate buffers and nucleic acid precursors; b) subjecting said nucleotide amplification reaction mixture to at least one thermocycle such that a first primer extension product is formed and cleaved at the ribonucleotide base releasing said first primer extension product; c) hybridizing said first primer extension product to a first DNA triggering template comprising a target sequence, a first primer extension product binding site at the 3′ terminus of said target sequence, and a contiguous second primer sequence which is conjoined to the 5′ end of said target sequence by a ribonucleotide base; d) subjecting said nucleotide amplification reaction mixture to at least one thermocycle such that a target amplification product is formed and said first DNA triggering template is cleaved at the ribonucleotide base releasing said second primer sequence with a ribonucleotide base at the 3′ terminus; e) hybridizing said second primer sequence to a second DNA triggering template which contains a second primer sequence binding site at the 3′ terminus; and f) subjecting said nucleotide amplification reaction mixture to at least one thermocycle such that a third primer extension product is formed and cleaved at the ribonucleotide base releasing said third primer extension product, wherein said third primer extension product has the same nucleotide sequence as the first primer extension product.