Method for constructing nucleic acid molecule
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United States of America Patent
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Feb 26, 2016
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Feb 26, 2015
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The present invention provides a method for constructing a single-stranded nucleic acid molecule for nucleic acid sequencing by means of a nanopore sequencer, said method including: a step in which at least one hairpin primer including a single-stranded region on the 3′ side and a pair of primers are used to synthesize a complementary strand of template DNA that includes the target sequence; and a step in which the synthesized complementary strand forms a hairpin structure inside a molecule and a template extension reaction is carried out. The obtained nucleic acid molecule includes both the target sequence and the complementary strand thereof in the sequence. Single strand construction enables analysis by nanopore sequencing, and the sequence of only the target nucleic acid, which does not include information of the complementary strand, is repeatedly analyzed, thus enabling analysis to be conducted with greater precision by addressing the problem of sequence errors.
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