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Application of magnetic nanoparticles in nucleic acid extraction and preparation method thereof

A magnetic nanoparticle and nanoparticle technology, applied in the field of nanomaterials, can solve problems such as low adsorption efficiency, and achieve the effects of reducing preparation cost, simplifying preparation process, and high adsorption

Active Publication Date: 2019-01-11
SUZHOU BAIYUAN GENT CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Therefore, the technical problem to be solved in the present invention is to overcome the defect that the magnetic nanoparticles used for extracting nucleic acid in the prior art have low adsorption efficiency.

Method used

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  • Application of magnetic nanoparticles in nucleic acid extraction and preparation method thereof
  • Application of magnetic nanoparticles in nucleic acid extraction and preparation method thereof
  • Application of magnetic nanoparticles in nucleic acid extraction and preparation method thereof

Examples

Experimental program
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Embodiment 1

[0058] This embodiment provides a magnetic nanoparticle (PDA@Fe 3 o 4 ) preparation method, comprising the following steps:

[0059] (1) with FeCl 3 ·6H 2 O as raw material, magnetic Fe synthesized by hydrothermal method 3 o 4 Nanoparticles

[0060] FeCl 3 ·6H 2 O (1.35 g) was dissolved in ethylene glycol (40 mL) and a clear solution was formed by sonication. Then, sodium acetate (3.6 g) and polyethylene glycol 10000 (1.0 g) were added to the solution, and the mixture was vigorously stirred until a homogeneous dark yellow solution was obtained. The solution was put into a polytetrafluoroethylene high pressure reactor and heated at 200° C. for 48 hours. The product was washed several times with ethanol and water, and dried under a nitrogen atmosphere at 60 °C for 10 h to obtain carboxyl-modified magnetic Fe 3 o 4 Nanoparticles, spare;

[0061] (2) Dopamine hydrochloride (200 mg) was mixed with 0.12 g of Tris-HCl buffer (10 mM, pH 8.5) and 100 ml of water to prepare ...

Embodiment 2

[0076] This embodiment provides a nucleic acid extraction kit, including nucleic acid binding solution and magnetic nanoparticles:

[0077] The nucleic acid binding solution includes 20% (w / v) polyethylene glycol, 4mol / L sodium chloride, and the pH of the nucleic acid binding solution is 2;

[0078] The magnetic nanoparticles are the magnetic nanoparticles prepared in Example 1 (PDA@Fe 3 o 4 ).

Embodiment 3

[0080] This embodiment provides a nucleic acid extraction method, using the nucleic acid extraction kit provided in Example 2, the extraction method includes the following steps:

[0081] (1) Cleavage of DNA in human whole blood samples

[0082] ①Put 300μl EDTA-K2 (EDTA dipotassium) anticoagulated blood into a 1.5ml centrifuge tube with a micropipette. Afterwards, 300 μl of sterile deionized water was added to the tube to disrupt the erythrocyte membrane. After repeated mixing several times, the mixture was centrifuged at 12000rpm / min for 3 minutes, and the supernatant was discarded to obtain a precipitate;

[0083] ② Add buffer CL (300 μl) to the precipitate, and mix the mixture several times. The mixture was centrifuged at 12000 rpm / min for 1 minute, and the supernatant was discarded;

[0084] ③ Prepare proteinase K-buffer FG mixture (1:100) (g / V) and add it to the precipitate. Immediately invert the solution slightly several times until the precipitate is completely dis...

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Abstract

The invention discloses an application of magnetic nanoparticles in nucleic acid extraction, wherein the magnetic nanoparticles are magnetic Fe3O4 nanoparticles with surfaces coated with a polydopamine coating. The magnetic nanoparticles provided by the invention have superparamagnetism, high environmental stability and monodispersity, the polydopamine coating coating on the surfaces of the nanoparticles is rich in amino group and hydroxyl group, can realize high-efficiency adsorption for nucleic acid, improves the efficiency and quality for adsorbing nucleic acid of existing magnetic nanoparticles, can realize high-efficiency rapid extraction of nucleic acid, does not need blood processing when being applied to human whole blood extraction, and thus is applicable to clinical disease genediagnosis and treatment. The preparation method of the magnetic nanoparticles disclosed by the invention effectively simplifies the preparation process of the magnetic nanoparticles used as a nucleicacid adsorbent, lowers the preparation process and is suitable for preparation and application of magnetic nanoparticles.

Description

technical field [0001] The invention belongs to the technical field of nanomaterials, and in particular relates to an application of magnetic nanoparticles in nucleic acid extraction and a preparation method thereof. Background technique [0002] Nucleic acid is the material basis for storing, replicating and transmitting genetic information, and is the key substance for a series of major life activities such as normal growth, development, reproduction, inheritance and variation of organisms. Therefore, the isolation and purification of nucleic acid is the primary problem to be solved in genetic engineering or protein engineering research. Meanwhile, the isolation of DNA is a crucial process in molecular biology and an essential step to initiate other downstream activities such as sequencing, amplification, hybridization, ligation, cloning, and biodetection. [0003] At present, according to the different nucleic acid extraction methods, nucleic acid extraction techniques c...

Claims

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Application Information

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IPC IPC(8): C12N15/10C01G49/08B82Y40/00B82Y30/00
CPCC12N15/1013B82Y30/00B82Y40/00C01G49/08C01P2004/62C01P2006/42C01P2002/72C01P2002/82C01P2004/04Y02A50/30
Inventor 尤崇革李琳张敏张俊平
Owner SUZHOU BAIYUAN GENT CO LTD
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