Methods for DNA sequencing with thermus aquaticus DNA polymerase
First Claim
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1. In a method for determining a nucleotide sequence for a nucleic acid segment by a dideoxynucleoside- 5'"'"'-triphosphate chain termination procedure, wherein said sequence is determined by extending an oligonucleotide primer in a templatedependent manner in the presence of an agent for polymerization, four dideoxynucleoside- 5'"'"'-triphosphates (dNTPs), and a dideoxynucleoside- 5'"'"'-triphosphate (ddNTP), the improvement comprising extending said primer in the presence of the agent for polymerization that is Thermus aquaticus DNA polymerase.
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Abstract
Dideoxynucleotide DNA sequencing methods can be dramatically improved by utilizing the DNA polymerase from Thermus aquaticus to catalyze the primer extension reactions.
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Citations
31 Claims
- 1. In a method for determining a nucleotide sequence for a nucleic acid segment by a dideoxynucleoside- 5'"'"'-triphosphate chain termination procedure, wherein said sequence is determined by extending an oligonucleotide primer in a templatedependent manner in the presence of an agent for polymerization, four dideoxynucleoside- 5'"'"'-triphosphates (dNTPs), and a dideoxynucleoside- 5'"'"'-triphosphate (ddNTP), the improvement comprising extending said primer in the presence of the agent for polymerization that is Thermus aquaticus DNA polymerase.
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26. A kit for determining a nucleotide sequence for a nucleic acid segment by a dideoxynucleoside-5'"'"'-triphosphate chain termination procedure, which kit comprises
(a) a primer for extending, in a template-dependent manner, a nucleic acid comprising a sequence complementary to said nucleic acid segment; -
(b) four deoxyribonucleoside-5'"'"'-triphosphates (dNTPs); (c) four dideoxyribonucleoside-5'"'"'-triphosphates (ddNTPs); and (d) Taq polymerase. - View Dependent Claims (27, 28, 29, 30, 31)
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Specification