Systems and methods for genetic and biological analysis
First Claim
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1. A method for nucleic acid sequencing, comprising:
- (a) providing an array comprising a plurality of features in optical communication with a plurality of optical sensors, wherein said array is integrated with said plurality of optical sensors, wherein a feature of said plurality of features retains or immobilizes a plurality of nucleic acid molecules having sequences that are substantially identical to a nucleic acid sequence of a template nucleic acid molecule, and wherein an optical sensor of said plurality of optical sensors is in optical communication with said feature;
(b) contacting said plurality of nucleic acid molecules with nucleic acid bases in the presence of polymerizing enzymes under conditions sufficient to incorporate said nucleic acid bases into growing strands complementary to said plurality of nucleic acid molecules, wherein said plurality of nucleic acid molecules is brought in contact with said nucleic acid bases without applying an electric field to confine or isolate said nucleic acid bases within said feature;
(c) using said optical sensor to detect one or more signals from said plurality of nucleic acid molecules indicative of said nucleic acid bases incorporated into said growing strands; and
(d) using said one or more signals detected in (c) to identify said nucleic acid bases, thereby generating a sequence corresponding to said nucleic acid sequence of said template nucleic acid molecule.
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Abstract
The invention relate to systems and methods for sequencing polynucleotides, as well as detecting reactions and binding events involving other biological molecules. The systems and methods may employ chamber-free devices and nanosensors to detect or characterize such reactions in high-throughput. Because the system in many embodiments is reusable, the system can be subject to more sophisticated and improved engineering, as compared to single use devices.
289 Citations
35 Claims
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1. A method for nucleic acid sequencing, comprising:
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(a) providing an array comprising a plurality of features in optical communication with a plurality of optical sensors, wherein said array is integrated with said plurality of optical sensors, wherein a feature of said plurality of features retains or immobilizes a plurality of nucleic acid molecules having sequences that are substantially identical to a nucleic acid sequence of a template nucleic acid molecule, and wherein an optical sensor of said plurality of optical sensors is in optical communication with said feature; (b) contacting said plurality of nucleic acid molecules with nucleic acid bases in the presence of polymerizing enzymes under conditions sufficient to incorporate said nucleic acid bases into growing strands complementary to said plurality of nucleic acid molecules, wherein said plurality of nucleic acid molecules is brought in contact with said nucleic acid bases without applying an electric field to confine or isolate said nucleic acid bases within said feature; (c) using said optical sensor to detect one or more signals from said plurality of nucleic acid molecules indicative of said nucleic acid bases incorporated into said growing strands; and (d) using said one or more signals detected in (c) to identify said nucleic acid bases, thereby generating a sequence corresponding to said nucleic acid sequence of said template nucleic acid molecule. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 29, 32, 33)
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22. A method for nucleic acid sequencing, comprising:
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(a) providing an array comprising a plurality of features in optical communication with a plurality of optical sensors, wherein said array is integrated with said plurality of optical sensors, wherein said plurality of features retains or immobilizes a plurality of nucleic acid molecules, wherein a feature of said plurality of features comprises a nucleic acid molecule of said plurality of nucleic acid molecules, and wherein an optical sensor of said plurality of optical sensors is in optical communication with said feature; (b) contacting said nucleic acid molecule with nucleic acid bases in the presence of a polymerizing enzyme under conditions sufficient to incorporate at least one of said nucleic acid bases into a growing strand complementary to said nucleic acid molecule; (c) using said optical sensor to detect one or more signals indicative of said at least one of said nucleic acid bases incorporated into said growing strand; (d) using said one or more signals detected in (c) to identify said at least one of said nucleic acid bases; and (e) monitoring and correcting for phase error introduced during incorporation of said at least one of said nucleic acid bases into said growing strand, thereby generating a sequence of said nucleic acid molecule. - View Dependent Claims (23, 24, 25, 26, 27, 28, 30, 31, 34, 35)
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Specification