Magnetic nano particle compound for nucleic acid separation and its preparation method and uses

A technology for separation of magnetic nanoparticles and nucleic acids, applied in the field of nanomaterials in the field of biotechnology, can solve problems such as small particle surface area, and achieve the effects of simple operation, easy control of shape and diameter

Inactive Publication Date: 2007-07-11
上海柏汇申生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] The technical problem to be solved in the present invention is to provide a kind of magnetic nanoparticle complex for nucleic acid separation and its preparation method and application, to overcome the prior art that only use the magnetic particle of common particle size to carry out nucleic acid separation, due to the particle surface area Small size, incapable of efficient separation of target nucleic acids in a short period of time

Method used

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  • Magnetic nano particle compound for nucleic acid separation and its preparation method and uses
  • Magnetic nano particle compound for nucleic acid separation and its preparation method and uses

Examples

Experimental program
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Effect test

Embodiment 1

[0049] Preparation of Magnetic Nanoparticles

[0050] Separately prepare FeSO with double distilled water 4 .7H 2 O and FeCl 3 .6H 2 O mixed solution and NaOH solution. In a mixed solution of iron salts, Fe 2+ The ion concentration is 0.1~0.2mol / L, Fe 3+ The concentration of ions is 0.1-0.3 mol / L, and the concentration of NaOH solution is 2-3 mol / L. Under vigorous stirring, the NaOH solution whose volume is half of the volume of the mixed salt solution was slowly added dropwise into the mixed salt solution. Aging the obtained solid precipitate at 40°C to 60°C for 12 hours, washing the precipitate several times with twice distilled water, filtering and drying at 40°C to 80°C for 24 hours, grinding it in an agate mortar That is the product.

Embodiment 2

[0052] Silica in γ-Fe 2 o 3 surface modification

[0053] Mix TritonX-100, n-hexanol, and cyclohexane uniformly in a ratio of 1:(1-3):(4-6) to form a transparent and stable microemulsion system. Put the above microemulsion system in ultrasonic treatment for 5-60 minutes, then add 0.01-1g of γ-Fe to it 2 o 3 (Magnetic nanoparticles), after ultrasonic treatment for 1-30 minutes, take out the upper layer and pour it into a three-necked flask, and stir for 30-60 minutes to make it uniform. Take 1-10mL of a certain concentration of concentrated ammonia water and dilute it with 1-20mL double-distilled water, slowly add it into the microemulsion that is constantly stirring, and keep stirring for 10-60 minutes to make the ammonia water evenly dispersed in the microemulsion. After 1 hour, 1-5 mL of tetraethyl orthosilicate (an inner-shell forming agent) was added dropwise to the microemulsion, while stirring continuously for 10 hours, and the temperature of the system was kept betw...

Embodiment 3

[0055] Surface Modification of Magnetic Nanoparticles with [N-(2-aminoethyl)-3-aminopropyltrimethoxysilane](AEAPS)

[0056] Take 15 mg of magnetic nanoparticles and add 10 to 100 mL of methanol and glycerol mixture (methanol: glycerol volume ratio is 2:3-2:1), and use ultrasonic treatment for 5 to 60 minutes; weigh 0.01 to 1g of AEAPS ([N-(2-aminoethyl)-3-aminopropyltrimethoxysilane]), treated with ultrasonic waves for 5-60 minutes; mix the two solutions evenly, and react at 10-95°C After 1-10 hours, the particles were taken out and washed three times with methanol, followed by vacuum drying at 100-300° C. to collect the particles (FSM).

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Abstract

The invention discloses a magnetic nanometer particle compound and making method and application to separate nucleic acid especially mRNA, which comprises the following three layers: (1) inner nuclear layer with magnetic nanometer particle with superparamagnetism, (2) biological inert outer layer to clad inner nuclear layer, (3) decorating layer with affinity or antibiotin on the surface of outer layer.

Description

technical field [0001] The invention belongs to the field of nanometer materials, and more specifically relates to nanomaterials applied in the field of biotechnology. Background technique [0002] Nucleic acid extraction, especially mRNA extraction technology is not only an important part of molecular biology technology, but also an important basis for functional genomics research technology. With the advent of the post-genome era, seeking an effective method for extracting nucleic acid, especially mRNA, has become an urgent problem to be solved. mRNA is messenger ribonucleic acid (messenger RNA). It is a messenger, or template, that exists in biological cells to transfer genetic information from DNA to functional proteins. It has become a research hotspot to elucidate the function of genes and the regulation mechanism of gene expression by studying gene expression profiling analysis. However, one of the biggest challenges in gene expression profiling is the extraction a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H1/06C07H21/00C07H21/02
Inventor 沈鹤柏徐文涛周海清
Owner 上海柏汇申生物科技有限公司
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