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Abstract

This invention relates to a method for selectively and efficiently synthesizing one of the sense or antisense strands of double-stranded nucleic acid. The method for synthesizing single-stranded nucleic acid according to this invention comprises the following steps of: 1) cleaving, with a restriction enzyme, double-stranded DNA having a restriction enzyme recognition sequence at a portion closer to the 5' side than the target sequence in such a manner that a) a fragment having an overhanging 3' terminus is formed, and b) base sequences of the single-stranded regions of the fragments after the cleavage differ from each other; 2) to the single-stranded region of the DNA fragment that was cleaved with the restriction enzyme, annealing a primer having a base sequence complementary to the region on at least its 3' terminus; and 3) synthesizing a nucleic acid by a strand displacement-type polymerase starting from the 3' terminus of the primer. Further, a single-stranded nucleic acid can be more efficiently synthesized by amplifying the double-stranded DNA having the restriction enzyme recognition sequence by the LAMP reaction.

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First Claim