Method for preparing magnetic silica microspheres with surfaces modified by cations

A technology of silica and surface modification, which is applied in the preparation of silica, silica, and test samples, etc., can solve the problems of nucleic acid loss, time-consuming and labor-intensive nucleic acid purification process, etc., and achieve large amount of nucleic acid, simple operation, and design reasonable effect

Active Publication Date: 2010-06-09
XIAN GOLDMAG NANOBIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, this nucleic acid purification process is time-consuming and laborious, and causes loss of nucleic acid during ethanol precipitation

Method used

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  • Method for preparing magnetic silica microspheres with surfaces modified by cations
  • Method for preparing magnetic silica microspheres with surfaces modified by cations
  • Method for preparing magnetic silica microspheres with surfaces modified by cations

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1 Preparation of magnetic silica microspheres with surface-modified histidine

[0051] 1. Preparation of Magnetic Silica Microspheres

[0052] Take 7.2892g of cetyltrimethylammonium bromide and 109.3g of toluene into the four-necked flask, stir evenly with a stirrer at 1000 rpm, mix 0.329g of FeCl 2 4H 2 O with 0.987g FeCl 3 ·6H 2 Dissolve O in 9.4397g of secondary water to prepare an aqueous solution with a concentration of 140g / L, add dropwise to the above mixture at 4-5mL / h under nitrogen protection at 2-3mL / min, and continue stirring at 800 rpm for 4-6 hours , then add 2 mL of ammonia water with a mass concentration of 25%, and continue to stir at 800 rpm for 2 hours. In the mixture, iron ferric oxide nanoparticles are formed, slowly add 9.79 mL of tetraethyl orthosilicate, and stir at 800 rpm. Adjust the pH to 8, 1 hour after adding tetraethyl orthosilicate dropwise at 2-3mL / h, stop the nitrogen protection, stir at 800 rpm, age for 5 days at normal temp...

Embodiment 2

[0057] Example 2 Preparation of magnetic silica microspheres with surface-modified polyhistidine

[0058] 1. Preparation of Magnetic Silica Microspheres

[0059] Take 7.2892g of cetyltrimethylammonium bromide and 109.3g of toluene into the four-necked flask, stir evenly with a stirrer at 1000 rpm, mix 0.329g of FeCl 2 4H 2 O with 0.987g FeCl 3 ·6H 2 Dissolve O in 9.4397g of secondary water to prepare an aqueous solution with a concentration of 140g / L, add dropwise to the above mixture at 4-5mL / h under nitrogen protection at 2-3mL / min, and continue stirring at 800 rpm for 4-6 hours , then add 2 mL of ammonia water with a mass concentration of 25%, and continue to stir at 800 rpm for 2 hours. In the mixture, iron ferric oxide nanoparticles are formed, slowly add 9.79 mL of tetraethyl orthosilicate, and stir at 800 rpm. Adjust the pH to 8, 1 hour after adding tetraethyl orthosilicate dropwise at 2-3mL / h, stop the nitrogen protection, stir at 800 rpm, age for 5 days at normal ...

Embodiment 3

[0064] Embodiment 3 prepares the magnetic silica microsphere of surface modification imidazole

[0065] 1. Preparation of Magnetic Silica Microspheres

[0066] Take 7.2892g of cetyltrimethylammonium bromide and 109.3g of toluene into the four-necked flask, stir evenly with a stirrer at 1000 rpm, mix 0.329g of FeCl 2 4H 2 O with 0.987g FeCl 3 ·6H 2 Dissolve O in 9.4397g of secondary water to prepare an aqueous solution with a concentration of 140g / L, add dropwise to the above mixture at 4-5mL / h under nitrogen protection at 2-3mL / min, and continue stirring at 800 rpm for 4-6 hours , then add 2 mL of ammonia water with a mass concentration of 25%, and continue to stir at 800 rpm for 2 hours. In the mixture, iron ferric oxide nanoparticles are formed, slowly add 9.79 mL of tetraethyl orthosilicate, and stir at 800 rpm. Adjust the pH to 8, 1 hour after adding tetraethyl orthosilicate dropwise at 2-3mL / h, stop the nitrogen protection, stir at 800 rpm, age for 5 days at normal te...

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Abstract

The invention relates to a method for preparing magnetic silica microspheres with surfaces modified by cations, and the number of electric charges of the modified cations varies with pH of solution. Ethyl orthosilicate is hydrolyzed to generate the magnetic silica microspheres by a sol-gel method in the presence of Fe3O4 nanoparticles. Epoxy silane reagent of 3-Glycidoxypropyltrimethoxy silane (GPTMS) is adopted to activate magnetic silica microparticles, and the cationic modifying agent with the pKa value within 4 to 8 is used as functional group, therefore, the magnetic silica microspheres with surfaces modified by cations are prepared by covalent interaction. The method comprises the preparation of the magnetic silica microspheres, the epoxidation of the magnetic silica microspheres and the cationic modification on the surfaces of the magnetic silica microspheres. The magnetic microspheres prepared by the method are used for separating nucleic acid from biological samples according to the variation of the electric charges on the surfaces.

Description

technical field [0001] The invention belongs to the field of material synthesis, and relates to a preparation method of a material used for separating nucleic acids from biological samples, in particular to a preparation method of surface-modified cation magnetic silica microspheres. Background technique [0002] Biomagnetic separation technology is a high-efficiency separation technology developed on the basis of traditional magnetic separation technology, which is applied to organisms such as cells, bacteria, nucleic acids and proteins. It uses the characteristics of magnetic or magnetically labeled organisms to move directionally under the action of an external magnetic field to extract, enrich, separate and purify target organisms. Early work mainly focused on the development of preparation techniques for magnetic markers, including the adsorption of magnetic nanoparticles to polymer microspheres such as agarose, polyacrylamide, polyglutaraldehyde, and polyvinylaldehyde ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C01B33/18G01N1/28C12N15/10
Inventor 崔亚丽杨磊胡道道陈超
Owner XIAN GOLDMAG NANOBIOTECH
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