Primer for gene amplification, method for nucleic acid discrimination with the use of the same, and nucleic acid discrimination kit

摘要

PCT No. PCT/JP96/00151 Sec. 371 Date Nov. 24, 1997 Sec. 102(e) Date Nov. 24, 1997 PCT Filed Jan. 26, 1996 PCT Pub. No. WO96/23077 PCT Pub. Date Aug. 1, 1996A nucleic acid differentiation method utilizing complementary strand-exchange reaction in competitive hybridization can clearly differentiate a site of gene mutation or a difference between genes in a target nucleic acid even when the gene mutation or difference is located near the primer binding site of the target nucleic acid. The invention provides a method and assay kit for differentiating the identity between two nucleic acids by labeling a gene amplifying primer including a primer main region which is complementary to the target nucleic acid to be amplified and having added to the 5' end of the primer main region a sequence which is noncomplementary to the target nucleic acid; amplifying one of two nucleic acids to be determined for identity using the labeled primer, thereby obtaining a labeled DNA; mixing the labeled DNA with an unlabeled DNA prepared from the other nucleic acid to effect competitive hybridization; and measuring the degree of complementary strand-exchange that has occurred between the labeled DNA and the unlabeled DNA by utilizing the label.