DNA fragment used as attached to 5′ end of primer used in nucleic acid amplification reaction and use of DNA fragment

摘要

A method is provided which enables quick, convenient, inexpensive, and high sensitivity confirmation of nucleic acid amplification after a nucleic acid amplification reaction. The DNA fragment in accordance with the present invention is a single-stranded DNA fragment containing a hairpin structure which in turn contains a bulge, wherein the DNA fragment is used as being attached to the 5′ end of a primer used in nucleic acid amplification. The nucleic acid amplification confirmation method in accordance with the present invention quantifies a hairpin primer containing the DNA fragment at its 5′ end by using bulge-binding fluorescent molecules after carrying out PCR or like nucleic acid amplification reaction using the hairpin primer. SNPs are detected quickly and conveniently at low cost and with high sensitivity by applying the nucleic acid amplification confirmation method, for example, to allele specific PCR.

主权项

1. A hairpin primer being an isolated DNA fragment for a polymerase chain reaction (PCR), comprising:
a nucleic acid fragment having a hairpin structure; and a primer segment, wherein: the hairpin structure contains bulges; the bulges are cytosine bulges; said nucleic acid fragment is attached to a 5′ end of the primer segment; and the hairpin primer contains any one of base sequences according to SEQ ID NOs 1 to 5.