Systems and methods for nucleic acid sequencing
First Claim
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1. A method for sequencing a nucleic acid template, comprising:
- (a) contacting a nucleic acid template with a sensing fluid containing a population of nucleotides, wherein said nucleic acid template is hybridized to a primer that is coupled to a bead, which bead is positioned proximate to a sensor in a sensor array, wherein said sensor comprises at least two electrodes, wherein said sensing fluid has a sensing fluid bulk conductivity and a surface of said bead has a surface conductivity to provide a Dukhin number that is less than 1 such that (i) a conductivity measurement by said at least two electrodes with said bead positioned proximate to said sensor is substantially similar to (ii) another conductivity measurement by said at least two electrodes without said bead positioned proximate to said sensor;
(b) using said at least two electrodes of said sensor to detect a conductivity change within a Debye layer of said bead upon incorporation of at least one nucleotide of said population of nucleotides into a growing nucleic acid strand, which growing nucleic acid strand is derived from said primer and is complementary to said nucleic acid template, which conductivity change is detected based at least in part on an electrical current change through said Debye layer;
(c) washing said sensor array to remove unincorporated nucleotides of said population of nucleotides from said sensor array; and
(d) repeating (a)-(c) to obtain sequence information for said nucleic acid template.
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Abstract
Provided herein are systems and methods for processing and analyzing nucleic acids and other biomolecules. Methods may include processing nucleic acid molecules in an emulsion of droplets. Methods of analyzing nucleic acid molecules may include coupling nucleic acids to a bead or other support. Methods may include analysis of nucleic acid molecules using a redox mediator. In some cases, analysis of the nucleic acid molecule includes determining a nucleotide sequence of the nucleic acid molecule.
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Citations
10 Claims
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1. A method for sequencing a nucleic acid template, comprising:
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(a) contacting a nucleic acid template with a sensing fluid containing a population of nucleotides, wherein said nucleic acid template is hybridized to a primer that is coupled to a bead, which bead is positioned proximate to a sensor in a sensor array, wherein said sensor comprises at least two electrodes, wherein said sensing fluid has a sensing fluid bulk conductivity and a surface of said bead has a surface conductivity to provide a Dukhin number that is less than 1 such that (i) a conductivity measurement by said at least two electrodes with said bead positioned proximate to said sensor is substantially similar to (ii) another conductivity measurement by said at least two electrodes without said bead positioned proximate to said sensor; (b) using said at least two electrodes of said sensor to detect a conductivity change within a Debye layer of said bead upon incorporation of at least one nucleotide of said population of nucleotides into a growing nucleic acid strand, which growing nucleic acid strand is derived from said primer and is complementary to said nucleic acid template, which conductivity change is detected based at least in part on an electrical current change through said Debye layer; (c) washing said sensor array to remove unincorporated nucleotides of said population of nucleotides from said sensor array; and (d) repeating (a)-(c) to obtain sequence information for said nucleic acid template. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
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Specification