Sequencing by orthogonal synthesis
First Claim
1. A system for sequencing nucleic acid templates, the system comprising:
- (a) an array of sites, wherein each site comprises a first nucleic acid template and a second nucleic acid template, wherein the first nucleic acid template comprises a DNA primer bound to the first template, and wherein the second nucleic acid template comprises an RNA primer bound to the second template;
(b) a DNA polymerase species suitable for extending the DNA primer bound to the first template;
(c) an RNA polymerase suitable for extending the RNA primer bound to the second template;
(d) a set of deoxyribonucleotide analogs suitable for producing a DNA primer extension product extending from the DNA primer, wherein the extension product comprises a deoxyribonucleotide analog at each of the sites;
(e) a set of ribonucleotide analogs suitable for producing an RNA primer extension product extending from the RNA primer, wherein the extension product comprises a ribonucleotide analog at each of the sites; and
(f) a detector for detecting the DNA primer extension product and the RNA primer extension product.
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Abstract
A method for sequencing includes steps of (a) providing first and second nucleic acid templates, wherein the two templates have different sequences; (b) extending a first primer bound to the first template using a first polymerase species and a first set of nucleotide analogs; (c) extending a second primer bound to the second template using a second polymerase species and a second set of nucleotide analogs, wherein the first polymerase species is different from the second polymerase species and wherein the first set of nucleotide analog is different from the second set of nucleotide analog, (d) detecting the first and second primer extension products; and (e) repeating steps (b) through (d), thereby determining the different sequences of the first and second templates.
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Citations
22 Claims
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1. A system for sequencing nucleic acid templates, the system comprising:
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(a) an array of sites, wherein each site comprises a first nucleic acid template and a second nucleic acid template, wherein the first nucleic acid template comprises a DNA primer bound to the first template, and wherein the second nucleic acid template comprises an RNA primer bound to the second template; (b) a DNA polymerase species suitable for extending the DNA primer bound to the first template; (c) an RNA polymerase suitable for extending the RNA primer bound to the second template; (d) a set of deoxyribonucleotide analogs suitable for producing a DNA primer extension product extending from the DNA primer, wherein the extension product comprises a deoxyribonucleotide analog at each of the sites; (e) a set of ribonucleotide analogs suitable for producing an RNA primer extension product extending from the RNA primer, wherein the extension product comprises a ribonucleotide analog at each of the sites; and (f) a detector for detecting the DNA primer extension product and the RNA primer extension product. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21)
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22. An array for orthogonal sequencing of nucleic acid templates, the array comprising:
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(a) a plurality of sites, each site comprising; i) a first nucleic acid template hybridized to a DNA primer, wherein the primer comprises at least one deoxyribonucleotide analog incorporated by a DNA polymerase species suitable for extending the DNA primer bound to the first nucleic acid template; ii) a second nucleic acid template hybridized to a RNA primer, wherein the primer comprises at least one ribonucleotide analog incorporated by a RNA polymerase species suitable for extending the RNA primer bound to the second nucleic acid template; (b) optionally a DNA polymerase species suitable for extending the DNA primer bound to the first template; (c) optionally an RNA polymerase suitable for extending the RNA primer bound to the second template; (d) optionally a set of deoxyribonucleotide analogs suitable for producing a DNA primer extension product extending from the DNA primer, wherein the extension product comprises a deoxyribonucleotide analog at each of the sites; (e) optionally a set of ribonucleotide analogs suitable for producing an RNA primer extension product extending from the RNA primer, wherein the extension product comprises a ribonucleotide analog at each of the sites; and (f) optionally a detector for detecting the DNA primer extension product and the RNA primer extension product.
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Specification