A single-molecule nucleic acid sequencing device based on nano-counter electrodes

A molecular nucleic acid and electrode technology, applied in the field of single-molecule nucleic acid sequencing devices based on nano-counter electrodes, to achieve the effects of overcoming sequencing bias, low-cost sequencing, and increasing read length

Inactive Publication Date: 2017-05-31
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 problems existing in the current single-molecule DNA sequencing, by detecting the fluctuation of the conductivity of protein molecules such as DNA polymerase or RNA polymerase in the process of nucleic acid synthesis, inferring the base sequence on the single-stranded nucleic acid, and developing a low-efficiency Cost, Rapid Nucleic Acid Sequencing Devices

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  • A single-molecule nucleic acid sequencing device based on nano-counter electrodes

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Embodiment approach 1

[0044] Embodiment 1: Single-molecule nucleic acid sequencing device based on nano-counter electrodes

[0045] (1) Preparation of gold nano-counter electrode: The nano-electrode pair chip is prepared on a silicon chip by conventional photolithography process. The process includes oxidation, glue coating, electron beam exposure, metal deposition, PR cleaning, BOE etching, and again Glue coating and electron beam exposure were used to prepare nano-gold electrode pairs with exposed tips. The processing index is: the electrode width is 50 nanometers, the gap is 10 nanometers; the SiO2 layer is 150 nanometers.

[0046] (2) Insulation modification of the surface of the gold nanoelectrode: the surface of the gold electrode was modified with thioglycolic acid aqueous solution / hexadecane mercaptan ethanol solution, and an insulating layer was constructed on the electrode surface. Soak in solution for 5 minutes; soak in deionized water for 5 minutes; soak in absolute ethanol for short-t...

Embodiment approach 2

[0052] Embodiment 2: Single-molecule nucleic acid sequencing device based on nano-counter electrodes

[0053] (1) Preparation of platinum nano-counter electrode: The nano-electrode pair chip is prepared on a silicon chip using a conventional photolithography process. The process includes oxidation, glue coating, electron beam exposure, metal deposition, PR cleaning, BOE etching, and again Glue coating and electron beam exposure are used to prepare exposed nano-gold electrode pairs, and a platinum layer of several nanometers is further deposited on the nano-gold electrode pairs. The processing index is: the electrode width is 50 nanometers, the gap is 10 nanometers; the SiO2 layer is 150 nanometers.

[0054] (2) Insulation modification on the surface of platinum nanoelectrodes: The surface of gold electrodes was modified with thioglycolic acid aqueous solution / hexadecanethiol ethanol solution, and an insulating layer was constructed on the electrode surface. The specific proce...

Embodiment approach 3

[0060] Embodiment 3: Single-molecule nucleic acid sequencing device based on nano-counter electrodes

[0061] (1) Preparation of palladium nano-counter electrode: The nano-electrode pair chip is prepared on the silicon chip by conventional photolithography process. The process includes oxidation, glue coating, electron beam exposure, metal deposition, PR cleaning, BOE etching, and again Glue coating and electron beam exposure were used to prepare exposed gold nano-electrode pairs. A palladium layer of several nanometers is further deposited on the nano-gold electrode pair. The processing index is: the electrode width is 50 nanometers, the gap is 10 nanometers; the SiO2 layer is 150 nanometers.

[0062] (2) Insulation modification on the surface of the palladium nanoelectrode: the surface of the gold electrode was modified with thioglycolic acid aqueous solution / hexadecane mercaptan ethanol solution, and an insulating layer was constructed on the electrode surface. The specif...

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Abstract

The invention relates to a single-molecule nucleic acid sequencing device based on nanometer counter electrodes. The rapid sequencing of a single-molecular nucleic acid molecule is realized by the single-molecule nucleic acid sequencing device through detecting the conductive properties of nucleic acid polymerases. The single-molecule nucleic acid sequencing device based on the nanometer counter electrodes belongs to the third generation DNA sequencing devices and is characterized in that the nanometer counter electrodes are prepared, the nanometer counter electrodes are a pair of electrodes with a nanometer clearance, and the clearance between the electrodes is 5-50 nanometers; the same protein or protein complex is connected at the opposite parts of the tip ends of the two electrodes; the electrode couple is placed in a water solution, and an electric insulating layer covers the surfaces, in contact with the water solution of, the electrodes; one electrode is placed in the solution and used as a reference electrode for electrochemical detection. Single DNA polymerase molecule is fixed between one pair of nanometer electrodes, the variation of the electric conductivity of the single DNA polymerase molecule during synthesis sequencing is detected in the solution, and the detection of the four base sequences on a single-molecule DNA template is realized.

Description

technical field [0001] The present invention relates to a single-molecule nucleic acid sequencing device based on a nano-counter electrode. Generation 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 and 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 and challenges to life science and medicine. . The thorough deciphering of the genetic code of life will soon become possible, and the popularization of big data on genetic information will benefit the survival and health of every ordinary member of human society. [0003] The next-generation DNA sequencing technology that is currently in widespread use is the so-called second-generation sequencing ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/04G01N27/327
Inventor 陆祖宏李清宁李志宏胡广丁长松贾忠伟万成康玉麟丁韬力
Owner PEKING UNIV
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