Real-time redox sequencing chips
AssigneePacific Biosciences of California, Inc.
InventorsJonas Korlach, Stephen Turner, Lei Sun
Life of this patent
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Abstract
Real time redox sequencing methods, devices, and systems are described. Arrays of redox devices comprising one or two electrodes are used to provide sequence information about a template nucleic acid in a polymerase-template complex bound proximate to the electrode(s). A sequencing reaction mixture comprising nucleotide analogs comprising redox labels is introduced to the array of redox devices under conditions of polymerase mediated nucleic acid synthesis. The time sequence of incorporation of nucleotide analogs is determined by electrochemically identifying the redox labels of the nucleotide analogs that are incorporated into the growing strand.
Claims25
A chip for sequencing a plurality of single nucleic acid template molecules comprising:
- a substrate comprising a plurality of redox devices, each redox device comprising two nanoscale electrodes, one of which acts as an oxidizing nanoscale electrode and one of which acts as a reducing nanoscale electrode,
- a polymerase enzyme complex comprising a polymerase enzyme and a template nucleic acid, the complex attached to one of the nanoscale redox electrodes or to the substrate proximate to the nanoscale electrodes,
- a reaction mixture in contact with the substrate comprising a plurality of types of nucleotide analogs each comprising a different redox label attached to the phosphate portion of the nucleotide analog through a linker of a length such that the redox label can come into contact with the nanoscale electrode acting as an oxidizing nanoscale electrode and the nanoscale electrode acting as the reducing nanoscale electrode when the nucleotide analog is held in the polymerase enzyme active site under conditions whereby polymerase mediated nucleic acid synthesis occurs;
- a plurality of electrical connection sites for bringing current and voltage to the redox devices, and for receiving electrical signals from the devices.
A chip for sequencing a plurality of single nucleic acid template molecules comprising:
- a substrate comprising a plurality of redox devices, each redox device comprising one nanoscale electrode, which acts as an oxidizing nanoscale electrode and as a reducing nanoscale electrode,
- a polymerase enzyme complex comprising a polymerase enzyme and a template nucleic acid, the complex attached to the nanoscale redox electrode or to the substrate proximate to the nanoscale electrode,
- a reaction mixture in contact with the substrate comprising a plurality of types of nucleotide analogs each comprising a different redox label attached to the phosphate portion of the nucleotide analog through a linker of a length such that the redox label can come into contact with the nanoscale electrode when the nanoscale electrode is acting as an oxidizing nanoscale electrode and when the nanoscale electrode is acting as a reducing nanoscale electrode when the nucleotide analog is held in the polymerase enzyme active site under conditions whereby polymerase mediated nucleic acid synthesis occurs;
- a plurality of electrical connection sites for bringing current and voltage to the redox devices, and for receiving electrical signals from the devices.
Dependencies are inferred from the claim text.