In order to develop and provide a method for conveniently and efficiently producing a nucleic acid aptamer, in particular a DNA aptamer, having excellent bonding and specificity properties with a target substance, a production method for a nucleic acid aptamer comprises: a complex formation step in which a complex of a single-stranded nucleic acid and a target substance is formed by mixing, in a solution, a single-stranded nucleic acid library and the target substance； an immobilization step in which a solid support and the solution after the above-mentioned step are mixed, and the complex is immobilized on the solid support via the target substance and/or bonding elements adhered to the solid support； a recovery step in which the complex, immobilized on the solid support, is recovered from the solution； an amplification step in which the single-stranded nucleic acid is recovered from the complex, and then amplified by means of a nucleic acid amplification method； and a single-stranded nucleic acid preparation step in which double-stranded nucleic acid, obtained in the amplification step, is made to be single stranded, and then an in-molecule three-dimensional structure is formed.
