Devices and methods for sequencing nucleic acids

Patent metadata

Field	Value
Publication number	US-9618477-B2
Application number	US-201313967108-A
Country	US
Kind code	B2
Filing date	Aug 14, 2013
Priority date	Sep 30, 2010
Publication date	Apr 11, 2017
Grant date	Apr 11, 2017

Abstract

Methods and devices for sequencing nucleic acids are disclosed herein. Devices are also provided herein for measuring DNA with nano-pores sized to allow DNA to pass through the nano-pore. The capacitance can be measured for the DNA molecule passing through the nano-pore. The capacitance measurements can be correlated to determine the sequence of base pairs passing through the nano-pore to sequence the DNA.

First claim

What is claimed is:

1. A method for sequencing a nucleic acid, comprising:
providing a nucleic acid to be sequenced;
passing the nucleic acid to be sequenced through a nano-pore on a substrate, the nano-pore having a diameter of less than about 5 nm; and
the nano-pore measuring the capacitance across the nano-pore as the nucleic acid passes through the nano-pore using one or more capacitors,
wherein the substrate has two top electrodes that are substantially parallel to each other and on opposing sides of the nano-pore on the top surface of the substrate and two bottom electrodes that are substantially parallel to each other and on opposing sides of the nano-pore on the bottom surface of the substrate, and wherein the top electrodes are substantially perpendicular to the bottom electrodes and each intersection of the top and bottom electrodes forms a capacitor.

2. The method of claim 1 , further comprising correlating the capacitance measurements to known capacitance values for specific nucleic acids.

3. The method of claim 1 , wherein the nano-pore has a diameter of less than about 2 nm.

4. The method of claim 1 , wherein the nucleic acid is DNA.

5. The method of claim 4 , wherein the DNA comprises one of RNA, single stranded DNA, or double stranded DNA.

6. The method of claim 1 , wherein the nano-pore is sized to fit about 5 or less nucleotides within the volume of the nano-pore.

7. The method of claim 4 , wherein after the capacitance is measured the DNA in the pore shifts by one nucleotide and the capacitance is measured again.

8. The method of claim 4 , further comprising providing alternating current to move the DNA forwards and backwards through the nano-pore.

9. The method of claim 1 , wherein an electric field is provided to pull the nucleic acid through the pore.

10. The method of claim 4 , further comprising using an enzyme to control the DNA passing through the pore.

11. The method of claim 1 , wherein an enzyme is used to cleave portions of the nucleic acid.

12. The method of claim 1 , wherein four capacitors are formed by the top and bottom electrodes.

13. The method of claim 1 , wherein the nucleic acid to be sequenced has a known nucleotide sequence; further comprising correlating the capacitance values to the known nucleotide sequence of the nucleic acid.

14. The method of claim 13 , wherein the apparatus has four capacitors adjacent to the nano-pore.

15. The method of claim 13 , wherein one to five nucleotides can fit in an internal volume of the nano-pore.

16. The method of claim 15 , wherein the known nucleotide sequence contains all of the possible sequence permutations for the number of nucleotides that can fit in the internal volume of the nano-pore.

Assignees

• California Inst Of Techn

Inventors

• Walavalkar Sameer
• Scherer Axel
• Tombrello Thomas A
• Rajagopal Aditya
• Homyk Andrew P
• Garcia Erika

Classifications

• G01N33/48721

  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 title

• G01N27/447 Primary

  using electrophoresis · CPC title

• C12Q1/6869 Primary

  Methods for sequencing · CPC title

• B82Y30/00

  Nanotechnology for materials or surface science, e.g. nanocomposites · CPC title

• C12Q2563/116

  electrical properties of nucleic acids, e.g. impedance, conductivity or resistance · CPC title