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Method for extracting DNA by using magnetic nano composite material and its kit

A composite material and magnetic nanotechnology, applied in biochemical equipment and methods, microbial measurement/inspection, sugar derivatives, etc., can solve the problems of extraction and separation of magnetic nanocomposites, product failure, and weak magnetic particles. And other issues

Inactive Publication Date: 2004-10-13
上海东晟纳米科技发展有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the structure of polymer spheres is usually relatively loose, and the magnetic particles contained in them are easy to detach from the microspheres during long-term storage, thus affecting the performance of the product
Generally, the polymer microspheres obtained by emulsion polymerization are large in size, mostly in the micron range, which cannot meet the requirements of some application fields that require smaller particles.
Another type of magnetic particle coating material commonly used is polysaccharide molecules, such as dextran, chitosan, etc., but the coating layer formed by such molecules is more unstable, and its interaction with magnetic particles is weak, and the same There is a disadvantage that the magnetic particles are easy to fall off and cause product failure
[0011] Currently, there is no technology for extracting and isolating DNA using magnetic nanocomposites

Method used

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  • Method for extracting DNA by using magnetic nano composite material and its kit
  • Method for extracting DNA by using magnetic nano composite material and its kit

Examples

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preparation example Construction

[0035] The preparation method of magnetic nanocomposite material of the present invention comprises steps:

[0036] (1) Preparation of Fe 3 o 4 sol

[0037] Preparation of Fe with a particle size of 5-8 nm by co-precipitation at room temperature 3 o 4 Magnetic nanocomposite colloids.

[0038]A kind of preferred method is with water-soluble ferric salt, as ferric chloride, iron sulfate, iron nitrate etc. and divalent iron salt, as ferrous chloride, ferrous sulfate, ferrous nitrate etc. according to molar ratio 2: Prepare an aqueous solution with a total iron concentration of 2-4mol / L at a ratio of 1, pour it into ammonia water with a concentration of 5% under strong stirring, filter it after stirring for 10-60 minutes, and then wash Fe with high-purity water 3 o 4 Precipitate. Disperse the precipitate in 23% tetramethylammonium hydroxide solution and stir for 10-60 minutes to obtain dispersed Fe 3 o 4 Sol.

[0039] (2) coated silica

[0040] Coated silica is usually ...

Embodiment 1

[0063] Preparation of Magnetic Nanocomposites

[0064] Take 16.62 grams of FeCl 3 and 9.93 g FeCl 2 , dissolved in 250ml deoxygenated high-purity water, added to 5L 5% ammonia water under strong stirring, filtered after stirring for 30 minutes, washed the precipitate with high-purity water, then added 300ml of 23% tetramethylammonium hydroxide solution, stirred for 30 minutes, added High-purity water to a volume of 2.5L to obtain Fe 3 o 4 A sol with a concentration of 1.7g / L. Take the above sol 1.69L, and the concentration of pH=9.5 is 0.58% SiO 2 Mix 310ml of aqueous solution, adjust pH=10 with 0.5M HCl, stir for 3 hours, mix with solution containing 168ml of concentrated ammonia water (25%), 4720ml of ethanol, 4ml of tetraethyl orthosilicate, stir for 30 minutes, add 46.5ml of 1.7g / L Fe 3 o 4 Sol, after stirring for 6 hours, SiO with a particle size of 50 nm was obtained 2 coated magnetic particles. Continue to add 1ml of tetraethoxymercaptopropylsilane, stir and re...

Embodiment 2

[0066] Preparation of Magnetic Nanocomposites

[0067] The particle diameter that obtains according to embodiment 1 operation is the SiO of 50nm 2 After coating the magnetic particles, continue to add 4ml orthosilicate ethyl ester, after stirring for 4 hours, obtain the SiO with a particle size of 100nm 2 coated magnetic particles. Then 1 ml of epoxytetraethoxysilane was added, and the stirring was continued for 2 hours to obtain composite magnetic particles with surface-modified epoxy groups.

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Abstract

The present invention relates to a method for extracting DNA by using magnetic nano composite material and its kit. The described method includes the following steps: (a). making magnetic nano composite material by contacted with liquid sample containing DNA to form liquid mixture, in which the described magnetic nano composite material has superparamagnetic endonucleus, intermediate silicon dioxide covering layer and external shell, and its diameter is 50-300 nm; and (b). using magnetic field to separate the magnetic nano composite material in which DNA is adsorbed from the liquid mixture so as to can obtain the magnetic nano composite material in which DNA is adsorbed. Said invented method can simply and effectively extract out DNA from various samples.

Description

technical field [0001] The present invention relates to the field of biotechnology. More specifically, the present invention relates to a method and a kit for extracting DNA with a magnetic nanocomposite material. Background technique [0002] DNA extraction has a wide range of applications, such as extracting DNA from samples such as blood spots and semen spots for genetic fingerprint identification, and extracting DNA from various tissues to construct libraries, etc. DNA extraction is a key technology in molecular biology experiments. The quality of DNA extraction will directly affect the success of future experiments, such as PCR amplification, enzymatic experiments, etc. DNA extraction methods have been the constant pursuit of molecular biology workers. [0003] Several genomic DNA extraction methods commonly used at present have obvious deficiencies. [0004] Although the phenol-chloroform extraction method is suitable for most biological samples, the experimental op...

Claims

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Application Information

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IPC IPC(8): C07H21/04C12Q1/68
Inventor 王刚杨百全庄家骐江林杨文胜
Owner 上海东晟纳米科技发展有限公司
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