Preparation method of magnetic beads by extracting nucleic acid and application of method

A magnetic bead and nucleic acid technology, applied in DNA preparation, chemical instruments and methods, alkali metal oxides/hydroxides, etc., can solve the problems of easy aggregation of magnetic nanoparticles, low chemical stability, and insufficient surface hydroxyl groups. Achieve the effect of simple operation, fast and thorough solid-liquid separation, and good water solubility

Active Publication Date: 2014-07-09
滁州市方舟医疗科技有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the nano-Fe 3 o 4 Due to the effects of small size effect, magnetic dipole attraction, etc., magnetic nanoparticles are easy to agglomerate, and the chemical stability is not high, easy to be oxidized, and the surface hydroxyl group is insufficient, which makes it difficult to be directly applied to the biological field.

Method used

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  • Preparation method of magnetic beads by extracting nucleic acid and application of method
  • Preparation method of magnetic beads by extracting nucleic acid and application of method
  • Preparation method of magnetic beads by extracting nucleic acid and application of method

Examples

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Effect test

Embodiment 1

[0043] A preparation method for nucleic acid extraction magnetic beads, comprising the following specific steps:

[0044] S1, Fe 3 o 4 Preparation of nanoparticles

[0045] Weigh 300g of anhydrous ferric chloride and dissolve in 1L of ultrapure water, weigh 392g of ferrous ammonium sulfate hexahydrate and dissolve in 1L of ultrapure water, transfer the two solutions into a 5L reactor and stir at 200r / min for 30min , weigh 45g of polyethylene glycol-8000, dissolve it in 400ml of ultrapure water, and transfer it to the reaction kettle; pass argon to remove oxygen for 30 minutes, adjust the reaction temperature to 60°C, adjust the speed to 500r / min, and drop 25% Ammonia water until the pH value of the system is 11, heat up to 80°C, adjust the rotation speed to 100r / min and react for 3h; The concentration was 1% ammonia water, 5% NaCl and deionized water, and the precipitate was repeatedly washed 3 times until the washing liquid was neutral, and freeze-dried at -10°C for 48 hou...

Embodiment 2

[0054] A preparation method for nucleic acid extraction magnetic beads, comprising the following specific steps:

[0055] S1, Fe 3 o 4 Preparation of nanoparticles

[0056] Same as step S1 in embodiment one;

[0057] S2, preparation of nano silicon-based magnetic beads

[0058] Take by weighing the superparamagnetic Fe obtained in the above-mentioned steps S1 3 o 4 Disperse 100 g of nanoparticles in 3L of absolute ethanol, after ultrasonic treatment for 30min, add 1L of aqueous solution with a concentration of 50 g / L trisodium citrate; after stirring at 200r / min for 30min, add ammonia water with a mass concentration of 25% to the system The pH value of the system is 10; then add dropwise 100ml of ethanol solution with a volume ratio of 50% tetraethyl orthosilicate, control the dropping rate at 0.5ml / min, and the system temperature at 30°C, and react for 4 hours after the dropping; take out the reactant , Magnetic separation of surface silicon-modified Fe 3 o 4 Nanopart...

Embodiment 3

[0060] A preparation method for nucleic acid extraction magnetic beads, comprising the following specific steps:

[0061] S1, Fe 3 o 4 Preparation of nanoparticles

[0062] Same as step S1 in embodiment one;

[0063] S2, preparation of nano silicon-based magnetic beads

[0064] Take by weighing the superparamagnetic Fe obtained in the above-mentioned steps S1 3 o 4 Disperse 500 g of nanoparticles into 4 L of absolute ethanol, and after ultrasonic treatment for 30 min, add 1 L of 50 g / L sodium edetate aqueous solution; after stirring at 200 r / min for 30 min, add 25% Ammonia water until the pH value of the system is 10; then dropwise add 130ml of ethanol solution with a volume ratio of 50% tetraethyl orthosilicate, control the dropping rate at 0.5ml / min, and the system temperature at 50°C, and react for 5 hours after the dropping; take out reactant, magnetically separates the surface silicon-modified Fe 3 o 4 Nanoparticles, after washing 3 times with absolute ethanol, re...

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Abstract

The invention discloses a preparation method of magnetic beads by extracting nucleic acid. The surfaces of magnetic nanoparticles Fe3O4 are coated with silicon hydroxyl and then are dispersed into a 0.02% NaN3 aqueous solution; the prepared silicon-based magnetic bead is of a core-shell structure and comprises a Fe3O4 nanoparticle core and a SiO2 shell layer, wherein the outer surfaces of a plurality of Fe3O4 nano-particles are coated with the SiO2 shell layer; the silicon-based magnetic bead has the secondary particle size of 400nm-1 micron, measured by a nanometer laser particle analyzer, and has the saturation magnetization of 43.0-74.5emu / g after being subjected to ultrasonic dispersion for 1h; the prepared silicon-based magnetic bead can be widely applied to nucleic acid extraction and purification of biomaterials such as fresh animal tissues, animal tissue paraffin sections, plant leaves, seeds, whole blood, free serum, hairs, fingernails, stubs, saliva, bacteria or viruses. The method is simple in process flow, convenient to operate, and low in raw material cost and is suitable for being used in industrial mass production.

Description

technical field [0001] This aspect belongs to the technical field of magnetic material preparation, and in particular relates to a preparation method and application of nucleic acid extraction magnetic beads. Background technique [0002] Magnetic beads are a functionalized carrier coated with biologically active groups, which can be dispersed in the base liquid to form magnetic liquid materials. The biologically active groups can be coupled with various biologically active substances, and have the fluidity of liquid And the dual characteristics of solid magnetic granular materials. The magnetic beads can move and concentrate in a direction under the action of an external magnetic field, and can restore the original structure and state after the external magnetic field is removed, thus making the complex liquid-solid separation technology quick and easy. Targeted substances with high purity can be obtained by simple elution. At present, magnetic beads have been widely used...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/06B01J20/28B01J20/30C12N15/10
Inventor 杜德光冯凌云齐存森艾振江肖理慧冯爱青
Owner 滁州市方舟医疗科技有限责任公司
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