Patent ID: 6090543
Filing Date: 2000-07-18
Classification: C12N,C12Q,Y10S

Abstract:
A method of detecting the presence of a target nucleic acid molecule by detecting non-target cleavage products comprising:a) providing:i) a cleavage means,ii) a source of target nucleic acid, said target nucleic acid having a first region, a second region, a third region and a fourth region, wherein said first region is located downstream from said second region, said second region is contiguous to and downstream from said third region and said third region is located adjacent to and downstream from said fourth region;iii) a first oligonucleotide complementary to said fourth region of said target nucleic acid;iv) second and third oligonucleotides having 3' and 5' portions, wherein said 3' portion of said second oligonucleotide contains a sequence complementary to said third region of said target nucleic acid and wherein said 5' portion of said second oligonucleotide and said 3' portion of said third oligonucleotide each contain sequence fully complementary to said second region of said target nucleic acid, and wherein said 5' portion of said third oligonucleotide contains sequence complementary to said first region of said target nucleic acid;b) mixing said cleavage means, said target nucleic acid, said first oligonucleotide, said second oligonucleotide and said third oligonucleotide to create a reaction mixture under reaction conditions such that said first oligonucleotide is annealed to said fourth region of said target nucleic acid and wherein at least said 3' portion of said second oligonucleotide is annealed to said target nucleic acid and wherein at least said 5' portion of said third oligonucleotide is annealed to said target nucleic acid so as to create a cleavage structure and wherein cleavage of said cleavage structure occurs to generate non-target cleavage products, each non-target cleavage product having a 3'-hydroxyl group; andc) detecting said non-target cleavage products.