SHORT CYCLE METHODS FOR SEQUENCING POLYNUCLEOTIDES
First Claim
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1. A method for obtaining sequence information from a plurality of different target nucleic acids immobilized at discrete locations on a surface, wherein a plurality of the target nucleic acids comprise a common adaptor sequence and have a primer hybridized thereto;
- wherein the method comprises multiple cycles of the following steps;
(a) contacting the target nucleic acids with a solution that contains an enzyme and a detectably labeled nucleotide such that the enzyme catalyzes template-dependent addition of the labeled nucleotide to extend a plurality of the primers,wherein the labeled nucleotide is not a chain-terminating nucleotide, andwherein addition of the labeled nucleotide to a primer interferes with progression of the enzyme;
then(b) detecting addition of the labeled nucleotide to at least some of the primers, thereby indicating the presence of a complementary nucleotide in the target nucleic acid hybridized thereto.
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Abstract
The invention provides methods for sequencing a polynucleotide comprising stopping an extension cycle in a sequence by synthesis reaction before the reaction has run to near or full completion.
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Citations
6 Claims
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1. A method for obtaining sequence information from a plurality of different target nucleic acids immobilized at discrete locations on a surface, wherein a plurality of the target nucleic acids comprise a common adaptor sequence and have a primer hybridized thereto;
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wherein the method comprises multiple cycles of the following steps; (a) contacting the target nucleic acids with a solution that contains an enzyme and a detectably labeled nucleotide such that the enzyme catalyzes template-dependent addition of the labeled nucleotide to extend a plurality of the primers, wherein the labeled nucleotide is not a chain-terminating nucleotide, and wherein addition of the labeled nucleotide to a primer interferes with progression of the enzyme;
then(b) detecting addition of the labeled nucleotide to at least some of the primers, thereby indicating the presence of a complementary nucleotide in the target nucleic acid hybridized thereto. - View Dependent Claims (2, 3, 4, 5, 6)
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Specification