Systems and methods for genetic and biological analysis
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.
1. A method for nucleic acid sequencing, comprising:
(a) activating a chip comprising a flow cell, an array in fluidic communication with said flow cell, a plurality of optical sensors in optical communication with said array, and circuitry operatively coupled to said plurality of optical sensors, wherein said array retains a plurality of nucleic acid molecules having sequences that are substantially identical to a nucleic acid sequence of a template nucleic acid molecule in a manner such that said plurality of nucleic acid molecules is in optical communication with an optical sensor of said plurality of optical sensors;
(b) bringing said plurality of nucleic acid molecules in contact with nucleic acid bases in presence of polymerizing enzymes under conditions sufficient to incorporate at least a subset of said nucleic acid bases into growing strands complementary to said plurality of nucleic acid molecules;
(c) using said optical sensor to detect one or more signals 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 at least said subset of said nucleic acid bases to thereby generate a sequence corresponding to said nucleic acid sequence of said template nucleic acid molecule.
2. The method of claim 1 , wherein said circuitry is coupled to one or more computer processors external to said chip, and wherein at least one of (b)-(d) is implemented by said one or more computer processors.
3. The method of claim 2 , wherein (b)-(d) are implemented by said one or more computer processors.
4. The method of claim 1 , wherein more than one type of nucleic acid base is directed to said flow cell in a given nucleic acid sequencing cycle.
5. The method of claim 1 , wherein four types of nucleic acid bases are directed to said flow cell in a given nucleic acid sequence cycle.
6. The method of claim 1 , wherein more than one type of nucleic acid base is directed to said flow cell in sequential nucleic acid sequencing cycles.
7. The method of claim 1 , wherein said array retains said plurality of nucleic acid molecules within a Debye length of a surface of said array or a surface of a feature immobilized on said array during sequencing.
8. The method of claim 1 , wherein said plurality of optical sensors comprise complementary metal-oxide semiconductor (CMOS) sensors.
9. The method of claim 1 , wherein said plurality of optical sensors comprise charge-coupled device (CCD) sensors.
10. The method of claim 1 , wherein said optical sensor is used to detect said one or more signals after said nucleic acid bases are incorporated into said growing strands.
11. The method of claim 1 , wherein said optical sensor of said plurality of optical sensors is individually addressable.
12. The method of claim 1 , wherein said array comprises capture sequences that immobilize said plurality of nucleic acid molecules.
13. The method of claim 1 , further comprising, prior to (b), amplifying said template nucleic acid molecule to generate said plurality of nucleic acid molecules having sequences that are substantially identical to said nucleic acid sequence of said template nucleic acid molecule.
14. The method of claim 13 , wherein said template nucleic acid molecule is amplified by isothermal amplification.
15. The method of claim 1 , wherein said nucleic acid bases comprise optical labels, and wherein one or more of said optical labels generate said one or more signals detected in (c).
16. The method of claim 15 , wherein said optical labels comprise fluorophores, dyes, or quantum dots.
17. The method of claim 1 , wherein said one or more signals are fluorescence signals.
18. The method of claim 1 , wherein said nucleic acid bases comprise reversible terminators.
19. The method of claim 1 , wherein said array comprises a surface coating.
20. The method of claim 1 , wherein said flow cell directs said nucleic acid bases to said array.