Single-molecule capacitive nucleic acid sequencing with nanoscale electrode pairs
AssigneePacific Biosciences of California, Inc.
InventorStephen Turner
Life of this patent
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Abstract
Sequencing methods, devices, and systems are described. Arrays of nanoscale electronic elements comprising two electrodes separated by an insulating layer are used to provide sequence information about a template nucleic acid in a polymerase-template complex bound proximate to the insulating region. A sequencing reaction mixture comprising nucleotide analogs having impedance labels is introduced to the array of nanoscale electronic elements under conditions of polymerase mediated nucleic acid synthesis. The time sequence of incorporation of nucleotide analogs is determined by identifying the types of labels of the nucleotide analogs that are incorporated into the growing strand using measured impedance.
Claims19
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.
Dependencies are inferred from the claim text.