| 摘要 |
An object of the present invention is to provide an amplification method that inhibits amplification caused by erroneous annealing of a primer. Primers X1 and X2 are used in amplification of a target sequence including a target site showing a polymorphism. The primer X1 includes a sequence A1' and a sequence E1. The sequence A1' is complementary to a partial sequence A1 in a template nucleic acid, and has, in its 3' region, a base x1' complementary to a first base x1 at the target site in the 5' region of the sequence A1. The sequence E1 is noncomplementary to a partial sequence B1 adjacent to the 3' end of the partial sequence A1 in the template nucleic acid, and is bound to the 5' end of the partial sequence A1'. The primer X2 includes a sequence A2'. The sequence A2' is complementary to a partial sequence A2 in the template nucleic acid, and has, in its 3' region, a base x2' complementary to a second base x2 at the target site in the 5' region of the partial sequence A2. Each of the primers X1 and X2 has a base complementary to the target site in its 3' region. By these primers, when only a template in which the target site is the first base x1 is present, erroneous amplification of the target sequence having the second base x2 can be prevented. Thus, a false positive for the polymorphism of the second base x2 can be inhibited. |
