Template-dependent nucleic acid polymerization using oligonucleotide triphosphates building blocks
First Claim
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1. A method of amplifying a nucleic acid template, the method comprising the step of contacting the nucleic acid template with a nascent oligonucleotide-3′
- -OH, a template-dependent polymerase and at least one oligonucleotide triphosphate under conditions in which said nascent oligonucleotide-3 ′
-OH hybridizes with the nucleic acid template and said template-dependent polymerase is active in incorporating said at least one oligonucleotide triphosphate onto a growing 3′
-OH group of said nascent oligonucleotide-3′
-OH, thereby amplifying the nucleic acid template, wherein said template-dependent polymerase is a DNA polymerase.
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Abstract
A novel use of a template-dependent polymerase. The novel use is effected by employing the template-dependent polymerase for incorporating at least one oligonucleotide triphosphate onto a nascent oligonucleotide-3′-OH in a template-dependent manner.
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Citations
4 Claims
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1. A method of amplifying a nucleic acid template, the method comprising the step of contacting the nucleic acid template with a nascent oligonucleotide-3′
- -OH, a template-dependent polymerase and at least one oligonucleotide triphosphate under conditions in which said nascent oligonucleotide-3 ′
-OH hybridizes with the nucleic acid template and said template-dependent polymerase is active in incorporating said at least one oligonucleotide triphosphate onto a growing 3′
-OH group of said nascent oligonucleotide-3′
-OH, thereby amplifying the nucleic acid template, wherein said template-dependent polymerase is a DNA polymerase.
- -OH, a template-dependent polymerase and at least one oligonucleotide triphosphate under conditions in which said nascent oligonucleotide-3 ′
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2. A method of amplifying a nucleic acid template, the method comprising the step of contacting the nucleic acid template with a nascent oligonucleotide-3′
- -OH, a template-dependent polymerase and 4N oligonucleotide triphosphates each including N monomers, wherein N is an integer equal to 2 or 3, under conditions in which said nascent oligonucleotide-3′
-OH hybridizes with the nucleic acid template and said template-dependent polymerase is active in incorporating said at least one oligonucleotide triphosphate onto a growing 3′
-OH group of said nascent oligonucleotide-3′
-OH, thereby amplifying the nucleic acid template.
- -OH, a template-dependent polymerase and 4N oligonucleotide triphosphates each including N monomers, wherein N is an integer equal to 2 or 3, under conditions in which said nascent oligonucleotide-3′
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3. A method of exponentially amplifying a nucleic acid template, the method comprising the step of contacting the nucleic acid template with a pair of nascent oligonucleotides-3′
- -OH, said nascent oligonucleotides-3′
-OH being hybridizable with opposite strands of the nucleic acid template, a template-dependent polymerase and 4N oligonucleotide triphosphates each including N monomers, wherein N is an integer equal to 2 or 3, under conditions in which said nascent oligonucleotides-3′
-OH hybridize with said opposite strands of the nucleic acid template and said template-dependent polymerase is active in incorporating said at least one oligonucleotide triphosphate onto a growing 3′
-OH group of each of said nascent oligonucleotides-3′
-OH, thereby exponentially amplifying the nucleic acid template.
- -OH, said nascent oligonucleotides-3′
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4. A method of amplifying a nucleic acid template, the method comprising the step of contacting the nucleic acid template with a nascent oligonucleotide-3′
- -OH, a template-dependent polymerase, at least one oligonucleotide triphosphate and at least one nucleotide triphosphate, wherein said at least one oligonucleotide triphosphate and said at least one nucleotide triphosphate are selected such that at least one monomer of said at least one oligonucleotide triphosphate is absent from said at least one nucleotide triphosphate, under conditions in which said nascent oligonucleotide-3′
-OH hybridizes with the nucleic acid template and said template-dependent polymerase is active in incorporating said at least one oligonucleotide triphosphate onto a growing 3′
-OH group of said nascent oligonucleotide-3′
-OH, thereby amplifying the nucleic acid template, wherein said template-dependent polymerase is a DNA polymerase.
- -OH, a template-dependent polymerase, at least one oligonucleotide triphosphate and at least one nucleotide triphosphate, wherein said at least one oligonucleotide triphosphate and said at least one nucleotide triphosphate are selected such that at least one monomer of said at least one oligonucleotide triphosphate is absent from said at least one nucleotide triphosphate, under conditions in which said nascent oligonucleotide-3′
Specification
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Current AssigneeYeda Research and Development Co. Ltd. (Weizmann Institute of Science)
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Original AssigneeYeda Research and Development Co. Ltd. (Weizmann Institute of Science)
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InventorsKless, Hadar
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Primary Examiner(s)Benzion; Gary
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Assistant Examiner(s)Wilder; Cynthia B
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Application NumberUS12/003,476Publication NumberTime in Patent Office993 DaysField of SearchNoneUS Class Current435/91.2CPC Class CodesC12P 19/34 Polynucleotides, e.g. nucle...C12Q 1/686 Polymerase chain reaction [...C12Q 2527/125 Specific component of sampl...
