Methods and compositions for correcting misincorporation in a nucleic acid synthesis reaction
First Claim
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1. A method for sequencing a nucleic acid, the method comprising the steps of:
- (a) obtaining a nucleic acid duplex comprising a template and a primer hybridized thereto;
(b) exposing said duplex to a plurality of deoxynucleotide triphosphates, at least one of which is complementary to a nucleotide at a position in said template immediately adjacent to the 3′
-terminus of said primer, in the presence of a first polymerase lacking exonuclease activity and a second polymerase having exonuclease activity;
(c) identifying said deoxynucleotide triphosphate that is complementary to said nucleotide; and
(d) repeating steps (b) and (c).
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Abstract
The invention provides methods for correcting misincorporation of a nucleotide in a primer during a sequencing-by-synthesis reaction by using both a polymerase substantially lacking in exonuclease activity and an enzyme, preferably a polymerase, having exonuclease activity.
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Citations
17 Claims
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1. A method for sequencing a nucleic acid, the method comprising the steps of:
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(a) obtaining a nucleic acid duplex comprising a template and a primer hybridized thereto;
(b) exposing said duplex to a plurality of deoxynucleotide triphosphates, at least one of which is complementary to a nucleotide at a position in said template immediately adjacent to the 3′
-terminus of said primer, in the presence of a first polymerase lacking exonuclease activity and a second polymerase having exonuclease activity;
(c) identifying said deoxynucleotide triphosphate that is complementary to said nucleotide; and
(d) repeating steps (b) and (c). - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17)
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Specification