Method for separating impurities in solution by chemically modified solid nanopore arrays

A nanopore array and chemical modification technology, which is applied in the field of chemically modified solid-state nanopore arrays to separate impurities in solutions, can solve problems such as limited separation, difficult processing, and decreased accuracy

Inactive Publication Date: 2012-04-25
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These technologies have their own scope of application, but their main disadvantages are: 1) the separation process itself will introduce new impurities; 2) the degree of separation is limited, and it is impossible to achieve absolute separation of single-molecule impurities; these shortcomings may affect some specific applications. cause adverse consequences
For example, during DNA cloning and library preparation, if the impurity DNA molecule cannot be completely removed, the impurity will be amplified simultaneously with the target sequence in a large amount during the amplification process, and the purity and reliability of the prepared clone or library will decrease , the subsequent use of these samples, such as DNA sequencing, will cause a decrease in accuracy and difficulties in data post-processing, etc.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Implementation steps:

[0017] 1) A 4-inch polysilicon wafer is selected, and a low-stress self-supporting silicon nitride film with a thickness of 30 nm is prepared by a silicon process. Through focused ion beam (FIB) etching (15 seconds), or transmission electron microscope (TEM) etching (180-200 seconds), the density is 920 / μm 2 , a solid-state nanohole array with an average pore diameter of 30nm and laser-cut into a square nanohole chip with a side length of 5mm;

[0018] 2) Dip the silicon nitride nanoporous chip containing solid nanopores into a mixture containing 98% concentrated sulfuric acid and hydrogen peroxide solution (volume ratio 7:3) and heat to 95°C, hydrate for 30 minutes, so that the surface has a large amount of Silicon hydroxyl and silicon oxygen bonds;

[0019] 3) Treat the silicon nitride film with an organosilane molecule, such as: 50 μl of a methanol solution of 1.5% γ-aminopropylmethyldiethoxysilane (containing 0.1% Triton-X100 nonionic surfa...

Embodiment 2

[0026] Implementation steps:

[0027] 1) Using chemical vapor deposition (CVD) to grow a silicon nitride film with a thickness of 1 μm on a clean silicon wafer with a diameter of 2.5 inches and a thickness of 300 μm;

[0028] 2) On the silicon nitride film, a uniform Kodak was prepared by spin coating at a speed of 2000rpm TM KPR photoresist layer with a thickness of about 1 μm;

[0029] 3) Prepare a suspension of microspheres, that is, a very dilute water-methanol suspension (1 ng / ml) of gold nanoparticles with a diameter of 20 nanometers. 1 μl of the suspension was evenly coated on the photoresist by spin coating at a speed of 2000 rpm, and heated to 80° C. to rapidly evaporate the solvent. This method can obtain a loose distribution of nanoparticles on the photoresist and become a random mask with a density of about 560 / μm 2 ;

[0030] 4) After electron beam exposure, the unexposed photoresist is washed away;

[0031] 5) Use hydrofluoric acid to etch the exposed silico...

Embodiment 3

[0041] Implementation steps:

[0042] 1) The manufacturing process of the nanohole array is the same as the 1)-7) step of embodiment 2; wherein the average diameter of nano gold is 120nm, and all the other processes are constant, and the nanohole array with an average aperture of 180nm can be obtained, and its density is 8-15 / μm 2 .

[0043] 2) Treat the nanopore array with an aqueous solution of anti-HIV-1p24 monoclonal antibody (containing 0.9% sodium chloride and 0.1% TritonX-100) at a concentration of 1 μg / ml for 1 hour, and wash off the antibody molecules that are not firmly bound by a large amount of deionized water and surfactants.

[0044] 3) Add 50 μl concentration of 10 to the front of the nanopore array 3 µl 1 Add 50 μl of 0.9% sodium chloride solution to the reverse side of the HIV virus suspension, and apply a bias voltage of 100 mV for 60 minutes. The solution collected on the other side of the nanopore cannot detect HIV by PCR and agarose gel electrophoresi...

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PUM

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Abstract

The invention relates to a method for separating impurities in a solution by chemically modified solid nanopore arrays. The area of the solid nanopore arrays is not less than 250 square micrometers. The diameter of a single nanopore is 1-200nm. Pore density is not less than 1 pore/square micrometer. The inner wall of pores is modified with one component or a mixture of more than two components ata random ratio, wherein the components are selected from the group consisting of 6-28bp nucleic acid molecule, antigen molecule, 60-200kDa antibody molecule, peptide molecule containing 6-50 amino acid and silane molecule. When 1-50 microliters of a solution which contains impurity molecule at the concentration of 1-10 nmol.ml<-1> or impurity particles of less than 1microgram/ml passes through the solid nanopore arrays, the impurity molecule or particles are intercepted by the chemically modified nanopore arrays. The structural formula of the silane molecule is Y(CH2)nSiXmZ3-m, wherein n is 0-3.

Description

technical field [0001] The invention relates to a method for separating impurities in a solution by chemically modifying a solid nanopore array. Background technique [0002] Nanopore refers to a hole on the substrate with a thickness of nanometer to micrometer, with a diameter of 1 to hundreds of nanometers, and penetrates both sides of the substrate. Nanopores can be divided into two categories in terms of preparation methods. The natural nanopores formed by biomolecules are called biological nanopores, and the nanopores obtained by people using micro-nano processing technology are called solid-state nanopores. The application of biological nanopores originated in 1996, when Kasianowicz and his colleagues reported for the first time that single-stranded DNA or RNA passed through α-hemolysin nanopores under the action of an electric field, and obtained a blocking current (Blockade Current) when molecules passed through the pores. The phenomenon of blocking current amplitu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01D61/14B01D61/42B01D57/02
Inventor 谢骁刘丽萍吴宏文孙峰刘全俊陆祖宏
Owner SOUTHEAST UNIV
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