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Protein molecule electronic device based on nanopore structure

A technology of nanopore structure and electronic devices, which is applied in specific-purpose bioreactors/fermenters, biochemical instruments, biomass post-processing, etc., and can solve problems such as poor base sensitivity on nucleic acid chains

Inactive Publication Date: 2015-01-28
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The present invention aims at the problem of poor sensitivity of the through-hole current to the bases on the nucleic acid chain in the solid-state nanopore sequencing technology. By immobilizing single protein molecules such as DNA polymerase or RNA polymerase or exonuclease in the nanopore, the efficiency of the through-hole is improved. The ability of current to recognize bases on single-stranded nucleic acids, and the development of a low-cost, fast DNA sequencing device

Method used

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  • Protein molecule electronic device based on nanopore structure

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Effect test

Embodiment approach 1

[0044] Embodiment 1: DNA polymerase molecular electronic device based on nanopore structure

[0045] (1) Preparation of nanopore chips: A series of micro-nano processing methods such as photolithography, dry etching, wet etching, and reactive ion beam etching are used to prepare Si / SiO 2 / SiN suspended support membrane structure. Among them, SiO 2 As an insulating buffer layer, it plays the role of supporting the SiN film and reducing the capacitive effect in electrical measurement. Si mainly supports SiO 2 effect. Finally, a 300kV high-energy focused electron beam (TEM) is used to process a nanohole with a diameter of less than 10 nanometers in the center of the suspended membrane. Processing indicators: nanopore diameter: less than 10 nanometers; thickness: around 50 nanometers.

[0046] (2) The self-made nanopore surface was modified with amino groups. The specific processing procedure is as follows: plasma treatment of self-made nanopore chip for 3 minutes, immersi...

Embodiment approach 2

[0053] Embodiment 2: RNA polymerase molecular electronic device based on nanopore structure

[0054] (1) Preparation of nanopore chips: A series of micro-nano processing methods such as photolithography, dry etching, wet etching, and reactive ion beam etching are used to prepare Si / SiO 2 / SiN suspended support membrane structure. Among them, SiO 2 As an insulating buffer layer, it plays the role of supporting the SiN film and reducing the capacitive effect in electrical measurement. Si mainly supports SiO 2 effect. Finally, a 300kV high-energy focused electron beam (TEM) is used to process a nanohole with a diameter of less than 10 nanometers in the center of the suspended membrane. Processing indicators: nanopore diameter: less than 10 nanometers; thickness: around 50 nanometers.

[0055] (2) The self-made nanopore surface was modified with amino groups. The specific processing procedure is as follows: the self-made nanopore chip is subjected to plasma treatment for 3...

Embodiment approach 3

[0062] Embodiment 3: Urease molecular electronic device based on nanopore structure

[0063] (1) Preparation of nanopore chips: A series of micro-nano processing methods such as photolithography, dry etching, wet etching, and reactive ion beam etching are used to prepare Si / SiO 2 / SiN suspended support membrane structure. Among them, SiO 2 As an insulating buffer layer, it plays the role of supporting the SiN film and reducing the capacitive effect in electrical measurement. Si mainly supports SiO 2 effect. Finally, a 300kV high-energy focused electron beam (TEM) is used to process a nanohole with a diameter of less than 10 nanometers in the center of the suspended membrane. Processing indicators: nanopore diameter: less than 10 nanometers; thickness: around 50 nanometers.

[0064] (2) The self-made nanopore surface was modified with amino groups. The specific processing procedure is as follows: the self-made nanopore chip is subjected to plasma treatment for 3 minutes...

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Abstract

The invention relates to a protein molecule electronic device based on a nanopore structure. The protein molecule electronic device is a mono-enzyme biosensor which can be used for detecting a substrate molecule with extremely low concentration in a solution and sequencing the DNA (deoxyribonucleic acid) of the single molecule. The protein molecule electronic device belongs to a third-generation DNA sequencing apparatus. The protein molecule electronic device based on the nanopore structure is characterized in that 1) the solution is divided into two parts by virtue of a solid structure with a nanochannel, wherein the solutions at the two sides of the solid structure are communicated by virtue of the nanochannel; 2) a protein molecule or a protein complex molecule is fixed in the nanochannel; 3) electrodes are respectively put in the two solutions separated by virtue of the solid structure, and resistance of the protein molecule in the nanochannel is detected by detecting current between the electrodes; and 4) the dynamic features of protein conformation are determined according to the changes of a current value passing through the protein molecule, and the information of the protein-catalyzed reaction substrate is obtained.

Description

technical field [0001] The invention relates to a protein molecular electronic device based on a nanopore structure, which is a single-enzyme biosensor, which can be used to detect substrate molecules with extremely low concentration in solution, and can be used to sequence single-molecule DNA. The invention belongs to the third generation of DNA sequencing devices. Background technique [0002] DNA sequencing technology is one of the important milestones in the development of modern life sciences. In the past ten years, the emergence of low-cost, high-throughput DNA sequencing technology has caused the explosive growth of the largest and most core nucleic acid sequence information in the highly complex system of life, which has brought unprecedented opportunities for the development of life sciences and medicine. And challenges, the thorough deciphering of the genetic code of life will soon become possible, and the popularization of big data on genetic information will ben...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/34
CPCB01J19/0046B01J2219/00277B01J2219/00608B01J2219/00853
Inventor 陆祖宏李清宁赵清贾忠伟万成康玉麟丁韬力
Owner PEKING UNIV
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