Systems and methods for characterizing a molecule

• US 9,041,420 B2
• Filed: 05/13/2013
• Issued: 05/26/2015
• Est. Priority Date: 02/08/2010
• Status: Active Grant

• Current Assignee: Roche Sequencing Solutions, Inc. (Roche Holding AG)
• Original Assignee: Genia Technologies, Inc. (Roche Holding AG)
• Inventors: Chen, Roger J. A., Davis, Randy
• Primary Examiner(s): Nguyen, Vincent Q
• Application Number:
  US13/893,142
• Publication Number:
  US 20140014513A1
• Time in Patent Office: 743 Days
• Field of Search: 324/691, 324/717
• US Class Current: 324/717
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 2563/116 electrical properties of nucleic acids, e.g. impedance, conductivity or resistance
• CPC Class Codes: C12Q 2565/631 being a biochannel or pore
• CPC Class Codes: G01N 2015/0038 Investigating nanoparticles
• CPC Class Codes: G01N 27/3278 involving nanosized elements, e.g. nanogaps or nanoparticles nanopores G01N33/48721; magnetic beads G01N27/745
• 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
• CPC Class Codes: Y10T 29/49002 Electrical device making
• CPC Class Codes: Y10T 29/49117 Conductor or circuit manufacturing

Abstract

Techniques for characterizing a molecule are described herein. In one example, a portion of the molecule is trapped in a nanopore, a variable voltage is applied across the nanopore until the trapped portion of molecule is moved within the nanopore, and the molecule is characterized based on the electrical stimulus required to affect movement of at least a portion of the trapped portion of the molecule within the nanopore.

Claims

1. A method for nucleic acid sequencing, comprising:

(a) providing a chip comprising a membrane adjacent to an electrode;

(b) measuring a capacitance and/or resistance across said membrane; and

(c) based on said capacitance and/or resistance measured in (b), (i) applying a destruction stimulus if said membrane exhibits a capacitance less than about 5 fF/μ

m² and/or a resistance less than about 10 GΩ

across said membrane under an applied voltage of about 50 mV, or (ii) forming a nanopore in said membrane if said membrane exhibits a capacitance greater than about 5 fF/μ

m² and/or a resistance greater than about 10 GΩ

across said membrane under an applied voltage of about 50 mV.

2. The method of claim 1, wherein said destruction stimulus is applied if said membrane exhibits a capacitance less than about 5 fF/μ

m² and a resistance less than about 10 GΩ

across said membrane under an applied voltage of about 50 mV.

3. The method of claim 1, wherein said nanopore is formed in said membrane if said membrane exhibits a capacitance greater than about 5 fF/μ

m² and a resistance greater than about 10 GΩ

across said membrane under an applied voltage of about 50 mV.

4. The method of claim 1, further comprising forming said nanopore and directing a nucleic acid molecule to said nanopore.

5. The method of claim 1, wherein said electrode is coupled to an integrated circuit that processes a signal detected with the aid of said electrode.

6. The method of claim 1, wherein said membrane is in sensing proximity to an integrated circuit.

7. The method of claim 6, wherein said integrated circuit comprises a logic controller.

8. The method of claim 1, wherein said membrane is nanopore-less.

9. The method of claim 1, wherein said membrane is a lipid bilayer.

10. The method of claim 1, wherein said capacitance and/or resistance is measured using said electrode.

11. A system for nucleic acid sequencing, comprising:

(a) a chip comprising a membrane adjacent to an electrode; and

(b) a processor coupled to said chip, wherein said processor executes instructions stored on and/or provided by a memory coupled to said processor, which instructions implement a method comprising;

(1) measuring a capacitance and/or resistance across said membrane; and

(2) based on said capacitance and/or resistance measured in (1), (i) applying a destruction stimulus if said membrane exhibits a capacitance less than about 5 fF/μ

m² and/or a resistance less than about 10 GΩ

across said membrane under an applied voltage of about 50 mV, or (ii) forming a nanopore in said membrane if said membrane exhibits a capacitance greater than about 5 fF/μ

m² and/or a resistance greater than about 10 GΩ

across said membrane under an applied voltage of about 50 mV.

12. The system of claim 11, wherein said method comprises applying said destruction stimulus if said membrane exhibits a capacitance less than about 5 fF/μ

m² and a resistance less than about 10 GΩ

across said membrane under an applied voltage of about 50 mV.

13. The system of claim 11, wherein said method comprises forming said nanopore in said membrane if said membrane exhibits a capacitance greater than about 5 fF/μ

m² and a resistance greater than about 10 GΩ

across said membrane under an applied voltage of about 50 mV.

14. The system of claim 11, wherein said method further comprises forming said nanopore and directing a nucleic acid molecule to said nanopore.

15. The system of claim 11, wherein said electrode is coupled to an integrated circuit that processes a signal detected with the aid of said electrode.

16. The system of claim 11, wherein said membrane is in sensing proximity to an integrated circuit.

17. The system of claim 16, wherein said integrated circuit comprises a logic controller.

18. The system of claim 11, wherein said membrane is nanopore-less.

19. The system of claim 11, wherein said membrane is a lipid bilayer.

20. The system of claim 11, wherein said capacitance and/or resistance is measured using said electrode.