Method and reagent for sequencing
AssigneeHitachi, Ltd.
InventorsHideki Kambara, Guohua Zhou, Tomoharu Kajiyama, Shigeya Suzuki
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Abstract
The present invention provides: a method for nucleic acid analysis including the steps of subjecting a reaction solution containing a sample nucleic acid to complementary strand synthesis with the sample nucleic acid as a template, reacting pyrophosphate produced in the complementary strand synthesis with 30 to 800 μM AMP in the coexistence of pyruvate phosphate dikinase to produce ATP, performing luciferase reaction with the ATP as a reaction substrate, and detecting chemiluminescence generated in the luciferase reaction to determine the presence or absence of the complementary strand synthesis; and a kit therefor.
Claims15
A method for nucleic acid analysis comprising the steps of
- subjecting a reaction solution comprising a sample nucleic acid to complementary strand synthesis with the sample nucleic acid as a template,
- reacting pyrophosphate produced in the complementary strand synthesis with 30 to 800 μM AMP in the coexistence of pyruvate phosphate dikinase to produce ATP,
- performing luciferase reaction with the ATP as a reaction substrate, and
- detecting chemiluminescence generated in the luciferase reaction to determine the presence or absence of the complementary strand synthesis.
A kit for nucleic acid analysis comprising
- 1) DNA polymerase,
- 2) four nucleic acid substrates or derivatives thereof corresponding to bases A, G, T, and C,
- 3) AMP, phosphoenolpyruvate, and pyruvate phosphate dikinase, and
- 4) luciferase and luciferin,
- wherein the AMP has a concentration of 30 to 800 μM in a reaction solution and is intended for reaction with pyruvate phosphate dikinase in the reaction solution.
A kit for nucleic acid analysis comprising
- 1) DNA polymerase,
- 2) four nucleic acid substrates or derivatives thereof corresponding to bases A, G, T, and C,
- 3) AMP, phosphoenolpyruvate, and pyruvate phosphate dikinase, and
- 4) luciferase and luciferin,
- wherein the kit comprises an indication stating that the AMP is reacted at a concentration of 30 to 800 μM in a reaction solution with pyruvate phosphate dikinase.
Dependencies are inferred from the claim text.