Systems and methods for high efficiency electronic sequencing and detection
The present disclosure relates to systems and methods for high efficiency electronic sequencing of nucleic acids and molecular detection.
1. A method for nucleic acid sequencing, comprising:
(a) bringing a nucleic acid molecule in contact with a nucleotide or nucleotide analogue under conditions sufficient to incorporate said nucleotide or nucleotide analogue into a growing strand complementary to said nucleic acid molecule; and
(b) using an electronic sensor operatively coupled to said nucleic acid molecule to measure a steady state signal or change in steady state signal within a solution comprising said nucleic acid molecule to identify said nucleotide or said nucleotide analogue incorporated into said growing strand.
2. The method of claim 1 , wherein said nucleotide or nucleotide analogue is incorporated into said growing strand in presence of a reaction buffer, and wherein said change in steady state signal is measured in presence of a measurement buffer.
3. The method of claim 2 , wherein said reaction buffer and said measurement buffer are different buffers.
4. The method of claim 2 , wherein a conductivity of said measurement buffer is lower than a conductivity of said reaction buffer.
5. The method of claim 1 , further comprising repeating (a) and (b) to identify additional nucleotides or nucleotide analogues incorporated into said growing strand, thereby generating a sequence of said nucleic acid molecule.
6. The method of claim 1 , further comprising, subsequent to (b), bringing said nucleic acid molecule in contact with a nucleotide or nucleotide analogue of a different type than said nucleotide or nucleotide analogue.
7. The method of claim 1 , wherein (a) is performed by bringing said nucleic acid molecule in contact with a plurality of nucleotide or nucleotide analogues comprising said nucleotide analogue, wherein said plurality of nucleotide analogues includes nucleotides analogues of different types.
8. The method of claim 1 , wherein said electronic sensor comprises at least two electrodes that are electrically coupled to said nucleic acid molecule or a feature to which said nucleic acid molecule is attached.
9. The method of claim 1 , wherein said electronic sensor comprises at least two electrodes, and wherein said at least two electrodes are exposed to said solution comprising said nucleic acid molecule.
10. The method of claim 1 , wherein said nucleic acid molecule is coupled to a particle disposed adjacent to said electronic sensor.
11. The method of claim 10 , wherein said particle is magnetically immobilized adjacent to said electronic sensor.
12. The method of claim 10 , wherein said particle is electronically immobilized adjacent to said electronic sensor.
13. The method of claim 1 , wherein said change in said steady state signal is a differential measurement.
14. The method of claim 13 , wherein said differential measurement utilizes (i) at least one reference sensor, (ii) another nucleic acid molecule of known sequence as a reference, or (iii) cross-talk deconvolution.
15. The method of claim 1 , further comprising using cross-talk deconvolution to reduce noise from said change in steady state signal.
16. The method of claim 1 , wherein said nucleic acid molecule is among a plurality of nucleic acid molecules adjacent to a sensor array comprising said electronic sensor.