METHOD AND DEVICE FOR IDENTIFYING NUCLEOTIDE, AND METHOD AND DEVICE FOR DETERMINING NUCLEOTIDE SEQUENCE OF POLYNUCLEOTIDE
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Abstract
The present invention provides technology that uses current measurements to identify nucleotides and determine a nucleotide sequence in polynucleotides. The present invention calculates a modal value of a tunnel current that arises when a nucleotide or polynucleotide for analysis passes through between electrodes, and then employs the calculated modal value. The present invention accordingly enables direct rapid implementation to identify nucleotides and to determine a nucleotide sequence in a polynucleotide without marking.
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Citations
34 Claims
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1-10. -10. (canceled)
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11. A method for sequencing a polynucleotide, comprising:
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(a) providing a solid substrate comprising two electrodes that at least partially define a gap, wherein the two electrodes detect a current upon movement of the polynucleotide comprising a plurality of nucleotides through the gap; (b) subjecting the polynucleotide to movement through the gap; (c) measuring the current with time across the gap using the two electrodes during movement of the polynucleotide through the gap, wherein the current is measured at a sampling rate greater than about 1 kHz; (d) determining durations and current levels associated with the current measured in (c) to identify individual nucleotides of the plurality of nucleotides with time; and (e) determining a sequence of the polynucleotide based on an identification of the individual nucleotides of the plurality of nucleotides. - View Dependent Claims (12, 13, 14, 15, 16, 17, 18, 19, 20, 21)
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22. A system for sequencing a polynucleotide, comprising:
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a solid substrate comprising two electrodes that at least partially define a gap, wherein the two electrodes detect a current upon movement of the polynucleotide comprising a plurality of nucleotides through the gap; and a computing device that (i) measures the current with time across the gap using the two electrodes during movement of the polynucleotide through the gap, wherein the current is measured at a sampling rate greater than about 1 kHz, (ii) determines durations and current levels associated with the current to identify individual nucleotides of the plurality of nucleotides with time, and (iii) determines a sequence of the polynucleotide based on an identification of the individual nucleotides of the plurality of nucleotides. - View Dependent Claims (23, 24, 25, 26, 27)
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28. A method for sequencing a polynucleotide, comprising:
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(a) causing the polynucleotide to pass between electrodes, wherein the electrodes detect a current upon movement of the polynucleotide comprising a plurality of nucleotides between the electrodes; (b) using the electrodes to continuously measure current with time within a measurement time period; (c) identifying isolated current levels while continuously measuring the current with time, wherein at least a subset of the current levels are different upon comparison to a reference value(s); and (d) associating each of the isolated current levels with individual nucleotides of the plurality of nucleotides, thereby identifying a sequence of the polynucleotide. - View Dependent Claims (29, 30)
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31. A method for sequencing a polynucleotide, comprising:
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(a) causing the polynucleotide comprising a plurality of nucleotides to pass between electrodes separated by a distance comparable to a size of a given nucleotide of the plurality of nucleotides, wherein the electrodes detect a current upon movement of the polynucleotide between the electrodes; (b) measuring the current upon movement of the polynucleotide between the electrodes to yield a current distribution with time; (c) dividing the current distribution into a plurality of segments, wherein individual segments of the plurality of segments correspond to individual nucleotides of the plurality of nucleotides; and (d) comparing the plurality of segments to one or more reference values to identify the individual nucleotides of the plurality of nucleotides, thereby generating a sequence of the polynucleotide. - View Dependent Claims (32, 33, 34)
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Specification