Method of detecting primer extension reaction, method of discriminating base type, device for discriminating base type, device for detecting pyrophosphate, method of detecting nucleic acid and tip for introducing sample solution

摘要

Convenient techniques for discriminating the base type in a base sequence of a nucleic acid are provided. The technique includes the step (a) of preparing a sample solution containing a nucleic acid, a primer having a base sequence that includes a complementary binding region which complementarily binds to the nucleic acid, and a nucleotide; the step (b) of allowing the sample solution to stand under a condition to cause an extension reaction of the primer, and producing pyrophosphate when the extension reaction is caused; the step (c) of bringing the sample solution into contact with the front face of a H<+> hardly permeable membrane having H<+>-pyrophosphatase, which penetrates from front to back of the membrane, of which active site that hydrolyzes pyrophosphate being exposed to the front face; the step (d) of measuring the H<+> concentration of at least either one of the solution at the front face side of the H<+> hardly permeable membrane or the solution at the back face side of the H<+> hardly permeable membrane, in a state where the H<+>-pyrophosphatase is immersed in the solution; the step (e) of detecting the extension reaction on the basis of the result of measurement in the step (d) ; and the step (f) of discriminating the base type in the base sequence of the nucleic acid on the basis of the result of detection in the step (e).