DNA sequencing method using step by step reaction
First Claim
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1. A method for sequencing a template nucleic acid, comprising the steps of:
- adding an amount of dATP that dose not exceed 50 times the amount of a template DNA to a reaction vessel containing the template DNA for step by step complementary strand synthesis; and
subtracting the background luminescence intensity caused by dATP, which is a substrate for luciferase, from the measured luminescence intensity, and measuring the luminescence intensity involved in the complementary strand synthesis, wherein said luminescence intensity is caused by ATP, which is produced from pyrophosphoric acid generated through the complementary strand synthesis.
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Abstract
The present invention provides an inexpensive DNA sequencing method with high-sensitivity. The method of the present invention comprising the steps of, adding an given amount of dATP for step by step complementary strand synthesis and subtracting the background luminescence intensity caused by dATP from the measured luminescence intensity to obtain the luminescence intensity involved in complementary strand synthesis.
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10 Claims
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1. A method for sequencing a template nucleic acid, comprising the steps of:
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adding an amount of dATP that dose not exceed 50 times the amount of a template DNA to a reaction vessel containing the template DNA for step by step complementary strand synthesis; and
subtracting the background luminescence intensity caused by dATP, which is a substrate for luciferase, from the measured luminescence intensity, and measuring the luminescence intensity involved in the complementary strand synthesis, wherein said luminescence intensity is caused by ATP, which is produced from pyrophosphoric acid generated through the complementary strand synthesis. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
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