| 摘要 |
Disclosed is a method of polynucleic acid (e.g., DNA) sequencing which is based on the generation of pyrophosphate (PPi) that occurs when a complementary base is incorporated into a growing DNA strand being synthesized on a template. The method utilizes a cascade of enzymatic reactions catalyzed by hypoxanthine-phosphoribosyl transferase, xanthine oxidase, and peroxidase in addition to DNA polymerase and apyrase. The last chemical step in the cascade of reactions is the oxidation of a material such as an electrode or luminol by hydrogen peroxide. This generates a detectable electrical or optical signal. This method is independent of luciferase, does not require dATP analogue, and is intended to improve precision and sensitivity of DNA sequencing, and to lessen the unsynchronized polymerization.
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