Systems and methods for forming a nanopore in a lipid bilayer

Patent metadata

Field	Value
Publication number	US-10926486-B2
Application number	US-201816179791-A
Country	US
Kind code	B2
Filing date	Nov 2, 2018
Priority date	Feb 8, 2010
Publication date	Feb 23, 2021
Grant date	Feb 23, 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 nucleic acid base of a molecule, the method comprising:
recording a first electrical measurement of a first portion of a molecule as the molecule moves through a nanopore under an applied electrical stimulus, wherein the first portion of the molecule includes a nucleic acid base of interest;
determining whether the nanopore is blocked by the molecule such that the molecule cannot advance through the nanopore;
applying an electrical pulse with a higher amplitude than the applied electrical stimulus to drive the molecule through the nanopore if it is determined that the nanopore is blocked;
recording a second electrical measurement of a second portion of the molecule as the molecule moves further through the nanopore under the applied electrical stimulus, wherein the second portion of the molecule also includes the nucleic acid base of interest, 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 representing a sequence of a first plurality of bases and a second frame comprises the second electrical measurement representing a sequence of a second plurality of bases and wherein the first and second frames overlap; and
identifying the nucleic acid base of interest of the molecule by processing the first electrical measurement and the second electrical measurement together, wherein processing the first electrical measurement and the second electrical measurement together includes combining and deconvolving the measurements to resolve the nucleic acid base of interest in the overlapping frames.

2. A system for identifying a nucleic acid base of a molecule, the system comprising:
a sensing circuit coupled to a nanopore; and
a processor coupled to the sensing circuit, wherein the processor is programmed to:
control the sensing circuit to record a first electrical measurement of a first portion of a molecule as the molecule moves through a nanopore under an applied electrical stimulus, wherein the first portion of the molecule includes a nucleic acid base of interest;
determine whether the nanopore is blocked by the molecule such that the molecule cannot advance through the nanopore;
apply an electrical pulse with a higher amplitude than the applied electrical stimulus to drive the molecule through the nanopore if it is determined that the nanopore is blocked;
control the sensing circuit to record a second electrical measurement of a second portion of the molecule as the molecule moves further through the nanopore under the applied electrical stimulus, wherein the second portion of the molecule also includes the nucleic acid base of interest, 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 representing a sequence of a first plurality of bases and a second frame comprises the second electrical measurement representing a sequence of a second plurality of bases and wherein the first and second frames overlap; and
identify the nucleic acid base of interest of the molecule by processing the first electrical measurement and the second electrical measurement together, wherein processing the first electrical measurement and the second electrical measurement together includes combining and deconvolving the measurements to resolve the nucleic acid base of interest in the overlapping frames.

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

• Y10S977/942

  including protein logic element · CPC title

• B82Y10/00

  Nanotechnology for information processing, storage or transmission, e.g. quantum computing or single electron logic · CPC title

• C12Q1/6869

  Methods for sequencing · CPC title

• Y10S977/797

  Lipid particle · CPC title