Method and apparatus for detecting nucleotides

• US 9,228,976 B2
• Filed: 09/27/2013
• Issued: 01/05/2016
• Est. Priority Date: 04/24/2000
• Status: Expired due to Term

• Current Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Inventors: Sauer, Jon R., Van Zeghbroeck, Bart J.
• Primary Examiner(s): ARORA, AJAY
• Application Number:
  US14/039,142
• Publication Number:
  US 20140021048A1
• Time in Patent Office: 830 Days
• Field of Search: 438/49
• US Class Current: 1/1
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 1/6874 involving nucleic acid arrays, e.g. sequencing by hybridisation
• CPC Class Codes: G01N 27/447 using electrophoresis
• CPC Class Codes: G01N 27/44791 Microapparatus sample containers with integrated microfluidic structures B01L3/5027
• CPC Class Codes: G01N 33/48721 Investigating individual macromolecules, e.g. by translocation through nanopores Coulter counters in general G01N15/12; fabrication methods for nanoscale apertures B81B1/00; sequencing of nucleic acids C12Q1/68

Abstract

A system and method employing at least one semiconductor device, or an arrangement of insulating and metal layers, having at least one detecting region which can include, for example, a recess or opening therein, for detecting a charge representative of a component of a polymer, such as a nucleic acid strand proximate to the detecting region, and a method for manufacturing such a semiconductor device. The system and method can thus be used for sequencing individual nucleotides or bases of ribonucleic acid (RNA) or deoxyribonucleic acid (DNA). The semiconductor device includes at least two doped regions, such as two n-typed regions implanted in a p-typed semiconductor layer or two p-typed regions implanted in an n-typed semiconductor layer. The detecting region permits a current to pass between the two doped regions in response to the presence of the component of the polymer, such as a base of a DNA or RNA strand. The current has characteristics representative of the component of the polymer, such as characteristics representative of the detected base of the DNA or RNA strand.

Claims

1. A system comprising:

a substrate defining a pore extending through the substrate and including at least three voltage carrying regions disposed along the pore;

a plurality of interconnects, each interconnect of the plurality of interconnects uniquely connected to a voltage carrying region of the at least three voltage carrying regions;

one or more power supplies in electrical communication with the plurality of interconnects and defining potential differences between at least two pairs of voltage carrying regions selected from the at least three voltage carrying regions; and

one or more meters to measure a difference in electrical characteristics of each of the at least two pairs of voltage carrying regions, the electrical characteristic indicative of a base sequence of a nucleic acid.

2. The system of claim 1, wherein the electrical characteristic includes a current between the each pair of voltage carrying regions.

3. The system of claim 1, wherein the current is responsive to the base sequence of the nucleic acid.

4. The system of claim 1, further comprising two fluid volumes, the substrate separating the two fluid volumes, the fluid volumes in fluidic communication via the pore.

5. The system of claim 1, wherein the voltage carrying region includes a doped silicon region.

6. The system of claim 5, wherein the doped silicon region includes an n-type doped silicon region.

7. The system of claim 1, further comprising a plurality of variable conductivity regions disposed between the at least three voltage carrying regions.

8. The system of claim 1, wherein the pore has a diameter in a range of 1 nm to 10 nm.

9. The system of claim 1, further comprising a silicon dioxide layer disposed between the at least three voltage carrying regions and a fluid volume within the pore.

10. A system comprising:

a substrate defining a pore extending completely through the substrate and including at least three source or drain regions disposed at different positions separated longitudinally along the pore;

a plurality of interconnects, each interconnect of the plurality of interconnects connected to a unique source or drain region of the at least three source or drain regions;

one or more power supplies in electrical communication with the plurality of interconnects and defining potential differences between at least two pairs of source or drain regions selected from the at least three source or drain regions; and

one or more current meters to measure a difference in current passing between source or drain regions of each pair of the at least two pairs of source or drain regions, the current responsive to a base sequence of a nucleic acid.

11. The system of claim 10, further comprising two fluid volumes, the substrate separating the two fluid volumes, the fluid volumes in fluidic communication via the pore.

12. The system of claim 10, wherein the source or drain region includes a doped silicon region.

13. The system of claim 12, wherein the doped silicon region includes an n-type doped silicon region.

14. The system of claim 10, further comprising a plurality of variable conductivity regions disposed between the at least three source or drain regions.

15. The system of claim 10, wherein the pore has a diameter in a range of 1 nm to 10 nm.

16. The system of claim 10, further comprising a silicon dioxide layer disposed between the at least three source or drain regions and a fluid volume within the pore.

17. A method of nucleic acid sequencing comprising:

applying a nucleic acid to a system comprising;

a substrate defining a pore extending through the substrate and including at least three voltage carrying regions distributed along the pore, the nucleic acid passing through the pore;

a plurality of interconnects, each interconnect of the plurality of interconnects connected to a unique voltage carrying region of the at least three voltage carrying regions;

one or more power supplies in electrical communication with the plurality of interconnects and defining potential differences between at least two pairs of voltage carrying regions selected from the at least three voltage carrying regions; and

one or more meters to measure a difference in electrical characteristics of each of the at least two pairs of voltage carrying regions, the electrical characteristic responsive to a base sequence of the nucleic acid; and

measuring the electrical characteristic.

18. The method of claim 17, wherein the pore has a diameter in a range of 1 nm to 10 nm.

19. The method of claim 17, wherein the electrical characteristic is current.