Ultra-low amount DNA extraction kit and method based on silica magnetic compound particles

A technology of composite particles and silica, applied in the biological field, can solve the problems of high price and low success rate of ultra-low DNA extraction, and achieve the effect of simple method, removal of inhibitors and pollutants, and rapid extraction

Inactive Publication Date: 2010-09-01
长春市博坤生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is also a lack of professional kits for ultra-low-volume DNA extraction on the market. Ultra-low-volume DNA extraction mainly relies on general DNA extraction kits from foreign companies, such as Promega, Qiagen, etc. These kits are not only expensive , and the success rate for ultra-low-volume DNA extraction is low

Method used

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  • Ultra-low amount DNA extraction kit and method based on silica magnetic compound particles
  • Ultra-low amount DNA extraction kit and method based on silica magnetic compound particles

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Experimental program
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Effect test

Embodiment 1

[0028] Step 1: 1 μm Co 3 o 4 - Preparation of silica magnetic composite particles

[0029] 50nm Co was synthesized by the method reported in the literature 3 o 4 - Silica core-shell nanoparticles. After mixing 300ml of water and 700ml of methanol, add 15.0g of magnesium nitrate and 2.0g of polyethylene diamine, stir to dissolve, then add 12g of 50nm Co 3 o 4 -Silicon dioxide core-shell nanoparticles, reacted for 30 minutes at a stirring speed of 80 rpm. Then, 40 ml of concentrated ammonia water with a mass percent concentration of 25-28% was added to the solution, and 0.35 ml of tetraethyl orthosilicate was added dropwise at a stirring speed of 300 rpm for 8 hours of reaction. Use a permanent magnet to separate the solid from the reaction solution, and wash the resulting solid with high-purity water for 3 times to obtain Co 3 o 4 - Silica magnetic composite particles with an average size of 1 μm.

[0030] Step 2: Preparation of Ultra-Low Quantity DNA Extraction Kit

...

Embodiment 2

[0037] Preparation of an extraction kit for ultra-low DNA extraction using silica magnetic composite particles, including:

[0038] 0.5ml silicon dioxide magnetic composite particle solution, its component is the Co prepared by 25mg / ml embodiment 1 3 o 4 - Silica magnetic composite particles;

[0039] 100ml pH=4.5 adsorption solution, its composition is 6mol / L NaClO 4 Solution, Tris-HCl of 0.5mol / L, EDTA of 35mmol / l, Triton of 4.0% (w / v);

[0040] 100ml washing liquid, its component is the ethanol of 1mol / l sodium acetate, 75% volume percent;

[0041] 10ml eluate, its composition is 10mmol / l Tris-HCl, pH=8.3

Embodiment 3

[0043] Preparation of an extraction kit for ultra-low DNA extraction using silica magnetic composite particles, including:

[0044] 0.5ml silicon dioxide magnetic composite particle solution, its component is the Co prepared by 25mg / ml embodiment 1 3 o 4 - Silica magnetic composite particles;

[0045] 100ml pH=7.0 adsorption solution, its component is 1mol / L GuSCN solution, the Tris-HCl of 0.5mol / L, the EDTA of 5mmol / l, the Triton of 0.5% (w / v);

[0046] 100ml washing liquid, its component is the ethanol of 80% volume percentage;

[0047] 10ml eluate, its composition is 8mmol / l Tris-HCl, pH=8.3

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Abstract

The invention relates to an ultra-low amount DNA extraction kit which contains silica magnetic compound particles, an absorption mixture with pH of 4.5-7.0, a washing mixture, an eluent, wherein the absorption mixture contains 1-8mol/l salt solution, 0.5mol/l Tris-HCl, 5-50mmol/l EDTA and 0.5-5% w/v Triton, the washing mixture contains 0-2mol/l sodium acetate and ethanol with the volume percentage of 40-80% and the eluent contains 8-12mol/l Tris-HCl with the pH of 8.3. The invention realizes the ultra-low amount DNA extraction efficiently by using silica magnetic compound particles the surfaces of which have small-sized nanoscale bulges, has simple method, realizes quick extraction which can be completed within one hour and can effectively remove inhibitors and pollutants. The invention is applicable to medicine detection, pharmaceutical factories and molecular biology labs.

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 ultra-low-quantity DNA by using silica magnetic composite particles. Background technique [0002] Ultra-low DNA generally refers to the concentration of DNA in the liquid sample is less than or equal to 1ng / ml. Whether DNA can be successfully extracted from samples containing ultra-low amounts of DNA is a difficult problem in molecular biology experiments, and the development of ultra-low amounts of DNA extraction methods has always been one of the goals of molecular biologists. Ultra-low DNA extraction methods have a wide range of applications, such as extracting trace amounts of host DNA from biopharmaceutical fermentation broth for drug quality control and monitoring, extracting trace DNA from degraded or diluted biological samples for forensic identification, etc. [0003] Superparamagnetic materials ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 王延旭王刚戴洁董涛吴晓明
Owner 长春市博坤生物科技有限公司
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