Detection of genetic polymorphisms by allele specific amplification using modified primers and subsequent hybridization

摘要

This invention features a discrimination primer for amplifying a nucleic acid that includes a first base at a position suspected of a polymorphism and a second base immediately 3' to the first base. This primer includes (1) a first nucleotide, which is located at the 3' terminus of the primer and includes a base that is complementary to the first base; (2) a second nucleotide, which is located immediately 5' to the first nucleotide and includes a base that is not complementary to the second base; (3) a segment of nucleotides, which is located immediately 5' to the second nucleotide and is complementary to a part of the nucleic acid that is immediately 3' to the second base; and (4) a binding member of a specific binding pair covalently bonded to the 5' terminus of the segment. The first base of the nucleic acid can be mutant or wild-type.