Incorporation of modified nucleotides by archaeon DNA polymerases and related methods
First Claim
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1. A method for site-specific incorporation of derivatized dideoxynucleotides into DNA comprising reacting an archaeon Family B DNA polymerase, a primed DNA template and nucleotide solution containing at least one derivatized dideoxynucleotide to produce fragments of DNA with the derivatized dideoxynucleoside covalently attached to the 3′
- terminal residue wherein the derivatized dideoxynucleotide is incorporated more efficiently than the corresponding underivatized dideoxynucleotide.
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Abstract
The present invention is directed toward improving the efficiency of chain terminator incorporation by Family B archaeon DNA polymerases. Previously, the low efficiency of ddNTP, and more especially dye-labeled ddNTP, incorporation has limited the usefulness of this group of DNA polymerases in protocols requiring chain terminator incorporation.
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Citations
31 Claims
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1. A method for site-specific incorporation of derivatized dideoxynucleotides into DNA comprising reacting an archaeon Family B DNA polymerase, a primed DNA template and nucleotide solution containing at least one derivatized dideoxynucleotide to produce fragments of DNA with the derivatized dideoxynucleoside covalently attached to the 3′
- terminal residue wherein the derivatized dideoxynucleotide is incorporated more efficiently than the corresponding underivatized dideoxynucleotide.
- View Dependent Claims (4, 5, 6, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 27, 28, 29, 30, 31)
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2. A method for site-specific incorporation of acyclonucleotides into DNA comprising reacting an archaeon Family B DNA polymerase, a primed DNA template and nucleotide solution containing at least one acyclonucleotide to produce fragments of DNA with the acyclonucleotide covalently attached to the 3′
- terminal residue.
- View Dependent Claims (7, 19, 20, 21, 22, 23, 24, 25, 26)
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3. A method for site-specific incorporation of derivatized acyclonucleotides into DNA comprising reacting an archaeon Family B DNA polymerase, a primed DNA template and nucleotide solution containing at least one derivatized acyclonucleotide to produce fragments of DNA with the derivatized acyclonucleotide covalently attached to the 3′
- terminal residue.
Specification