Generation of modified polymerases for improved accuracy in single molecule sequencing
First Claim
1. A composition comprising a modified recombinant Φ
- 29-type DNA polymerase, which modified recombinant polymerase comprises an amino acid sequence that is at least 80% identical to SEQ ID NO;
1 and comprises an alanine substitution at position 253, wherein numbering of positions is relative to SEQ ID NO;
1, and which modified recombinant polymerase exhibits polymerase activity.
1 Assignment
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Accused Products
Abstract
Provided are compositions comprising modified recombinant polymerases that exhibit branching fractions that are less than the branching fractions of the polymerases from which they were derived, or branching fractions that are less than about 25% for a phosphate-labeled nucleotide analog. Also provided are compositions comprising modified recombinant polymerases that exhibit closed polymerase/DNA complexes with increased stability relative to the parental polymerases. Also provided are compositions comprising modified recombinant polymerases that exhibit decreased rate constants relative to the parental polymerases. Provided are methods for generating polymerases with the aforementioned phenotypes. Provided are methods of using such polymerases to make a DNA or to sequence a DNA template.
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Citations
12 Claims
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1. A composition comprising a modified recombinant Φ
- 29-type DNA polymerase, which modified recombinant polymerase comprises an amino acid sequence that is at least 80% identical to SEQ ID NO;
1 and comprises an alanine substitution at position 253, wherein numbering of positions is relative to SEQ ID NO;
1, and which modified recombinant polymerase exhibits polymerase activity. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12)
- 29-type DNA polymerase, which modified recombinant polymerase comprises an amino acid sequence that is at least 80% identical to SEQ ID NO;
Specification