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Single-molecule capacitive nucleic acid sequencing with nanoscale electrode pairs

  • US 9,868,987 B2
  • Filed: 10/21/2015
  • Issued: 01/16/2018
  • Est. Priority Date: 05/06/2013
  • Status: Active Grant
First Claim
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1. A method for single molecule nucleic acid sequencing comprising:

  • providing a substrate comprising an array of nanoscale capacitive devices, each capacitive device comprising two nanoscale electrodes separated by an insulating region, wherein a single polymerase enzyme complex comprising a single polymerase enzyme and a template nucleic acid is attached to the insulating region;

    exposing the substrate to a plurality of types of nucleotide analogs, each type comprising a different capacitive label attached to the phosphate portion of the nucleotide analog under conditions whereby polymerase mediated nucleic acid synthesis occurs, resulting in cleavage of the capacitive label and the growth of a nascent nucleic acid strand;

    applying a voltage across the two nanoscale electrodes in each device, whereby when a nucleotide analog resides in the active site of the enzyme, the capacitive label on the nucleotide analog produces a measurable change in the capacitance measured at the nanoscale electrodes, such change in capacitance occurring before the cleavage of the capacitive label;

    measuring the capacitance at the nanoscale electrodes over time, whereby the capacitance over time indicates an incorporation event and identifies the type of nucleotide analog by its capacitive label; and

    using the measured capacitance at the electrodes over time to determine a sequence of the template nucleic acid.

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