Patent ID: 8354232

Claim:
A method of target nucleic acid dependent amplification of a non-target sequence in a sample comprising; a) incubating a sample with stem oligonucleotides, upstream oligonucleotides, primers, dNTPs, and a polymerase under conditions such that, if a target nucleic acid is present: i) a 3′ target specific region of said stem oligonucleotides hybridizes to said target nucleic acid, and a stem region of said stem oligonucleotides remains available for hybridization to said upstream oligonucleotides, ii) a 5′ target specific region of said upstream oligonucleotides hybridizes to said target nucleic acid, and a stem specific region of said upstream oligonucleotides hybridizes to a portion of said stem region of said stem oligonucleotides, and iii) said polymerase extends a 3′ end of said upstream oligonucleotides using said stem region of said stem oligonucleotides as a template to generate extended upstream oligonucleotides that comprise an upstream oligonucleotide extended region; b) heating said sample to separate said extended upstream oligonucleotides from said stem oligonucleotides and said target nucleic acid; c) cooling said sample under conditions such that said primers hybridize to at least a portion of said upstream oligonucleotide extended region of said extended upstream oligonucleotides; and d) incubating said sample under conditions such that said primers are extended by said polymerase using said extended upstream oligonucleotides as templates such that stem amplicon sequences are generated.