Systems and methods for forming a nanopore in a lipid bilayer

Patent metadata

Field	Value
Publication number	US-11027502-B2
Application number	US-201816179800-A
Country	US
Kind code	B2
Filing date	Nov 2, 2018
Priority date	Feb 8, 2010
Publication date	Jun 8, 2021
Grant date	Jun 8, 2021

Abstract

Techniques for forming a nanopore in a lipid bilayer are described herein. In one example, an agitation stimulus level such as an electrical agitation stimulus is applied to a lipid bilayer wherein the agitation stimulus level tends to facilitate the formation of nanopores in the lipid bilayer. In some embodiments, a change in an electrical property of the lipid bilayer resulting from the formation of the nanopore in the lipid bilayer is detected, and a nanopore has formed in the lipid bilayer is determined based on the detected change in the lipid bilayer electrical property.

First claim

What is claimed is:

1. A method of identifying a base of a nucleic acid molecule, comprising:
recording a first electrical measurement indicative of the base of the nucleic acid molecule as a strand of the nucleic acid molecule moves through a nanopore under an applied electrical stimulus;
recording a second electrical measurement indicative of the base of the nucleic acid molecule wherein the second electrical measurement is a repeat electrical measurement; and
identifying the base based on the first electrical measurement wherein the repeat measurement is used to increase confidence in the identification, wherein the repeat electrical measurement corresponds to a different conformation of the nucleic acid molecule.

2. The method of claim 1 , wherein the repeat electrical measurement is made under substantially the same conditions as the first electrical measurement.

3. A method of identifying a base of a nucleic acid molecule, comprising:
recording a first electrical measurement indicative of the base of the nucleic acid molecule as a strand of the nucleic acid molecule moves through a nanopore under an applied electrical stimulus;
recording a second electrical measurement indicative of the base of the nucleic acid molecule wherein the second electrical measurement is a repeat electrical measurement; and
identifying the base based on the first electrical measurement wherein the repeat measurement is used to increase confidence in the identification, wherein the repeat electrical measurement corresponds to a different orientation of the nucleic acid molecule.

4. The method of claim 1 , wherein the repeat electrical measurement corresponds to the same base of the nucleic acid molecule.

5. The method of claim 1 , wherein the base of the nucleic acid molecule is part of a single strand of DNA that complements a second base.

6. A method of identifying a base of a nucleic acid molecule, comprising:
recording a first electrical measurement indicative of the base of the nucleic acid molecule as a first portion of the nucleic acid molecule moves through a nanopore under an applied electrical stimulus, wherein the first portion of the nucleic acid molecule includes the base;
recording a second electrical measurement indicative of the base of the nucleic acid molecule as a second portion of the nucleic acid molecule moves through the nanopore under the applied electrical stimulus, wherein the second portion of the nucleic acid molecule also includes the base, and wherein the second portion of the molecule is different from the first portion of the molecule;
combining and deconvolving the first electrical measurement and the second electrical measurement; and
identifying the base based on the first electrical measurement and the second electrical measurement.

7. A method of identifying a base of a nucleic acid molecule, comprising:
recording a first electrical measurement indicative of the base of the nucleic acid molecule as a first portion of the nucleic acid molecule moves through a nanopore under an applied electrical stimulus, wherein the first portion of the nucleic acid molecule includes the base;
recording a second electrical measurement indicative of the base of the nucleic acid molecule as a second portion of the nucleic acid molecule moves through the nanopore under the applied electrical stimulus, wherein the second portion of the nucleic acid molecule also includes the base, and wherein the second portion of the molecule is different from the first portion of the molecule, wherein a first frame comprises the first electrical measurement and a second frame comprises the second electrical measurement and wherein the first and second frames overlap; and
identifying the base based on the first electrical measurement and the second electrical measurement.

Assignees

• Roche Sequencing Solutions Inc

Inventors

• Chen Roger J A
• Davis Randy

Classifications

• B82Y5/00 Primary

  Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery · CPC title

• B82Y40/00

  Manufacture or treatment of nanostructures · CPC title

• Y10S977/797

  Lipid particle · CPC title

• 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

• Y10S977/90

  having step or means utilizing mechanical or thermal property, e.g. pressure, heat · CPC title