Preparation method of core-shell magnetic conductive polymer microspheres and application thereof

A conductive polymer and polymer technology, applied in the direction of DNA preparation, recombinant DNA technology, etc., can solve the problems of unfriendly environment and increased production cost

Inactive Publication Date: 2012-09-19
QILU UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The heterogeneous polymerization method can obtain high-quality core-shell magnetic composite microspheres, but cross-linking agents and / or surfactants need to be added during the preparation process; this not only increases the production cost, but is also environmentally unfriendly

Method used

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  • Preparation method of core-shell magnetic conductive polymer microspheres and application thereof
  • Preparation method of core-shell magnetic conductive polymer microspheres and application thereof
  • Preparation method of core-shell magnetic conductive polymer microspheres and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1: Core-shell Fe 3 O 4 / PANI composite microsphere preparation

[0056] Add 0.1g Fe into a 100mL three-necked flask 3 O 4 , 40mL 0.1mol / L hydrochloric acid, ultrasonic dispersion at room temperature for 5 minutes, sealed and placed in a 5 ℃ constant temperature box for 12 hours, magnetic separation to remove the aqueous hydrochloric acid solution, retaining the magnetic core; then add 15mL absolute ethanol, 0.1 g Re-distilled aniline, ultrasonically disperse for 5 minutes, and place it in a thermostat at 5°C for 12 hours.

[0057] Place the flask in an ultrasonic cleaner, and control the water temperature of the ultrasonic cleaner to 5-10°C. Dissolve 0.25mL of 12mol / L hydrochloric acid in 45mL of distilled water, then add 0.28g of ammonium persulfate to dissolve, slowly add the oxidant solution dropwise to control the dropping time to 0.5h; the reaction is maintained and stirred with the assistance of ultrasound for 3h.

[0058] Solid-liquid magnetic separation, water...

Embodiment 2

[0059] Example 2: Core-shell Fe 3 O 4 / PPy composite microsphere preparation

[0060] Replace the aniline in Example 1 with pyrrole, and use 0.40g FeCl for 0.28g ammonium persulfate 3 ·6H 2 O instead, other operations remain unchanged, you can get Fe 3 O 4 / PPy sample.

Embodiment 3

[0061] Example 3: Using Fe 3 O 4 / PANI composite microspheres to separate and purify E.coli DH5α plasmid DNA

[0062] Take 10mg de-doped Fe 3 O 4 / PANI magnetic ball is placed in a 1.5mL sterile centrifuge tube, sonicated with 1mL 70% (v / v) ethanol solution, sterilized twice, then transferred to another sterile centrifuge tube, and then added 40μL E.coli DH5α The crude plasmid DNA extract and 1mL Tris-HCl (10mmol / L, pH 7.8) buffer, shake well, and place it at room temperature for 20 minutes until the DNA is adsorbed on the surface of the magnetic ball. Collect the magnetic ball under a magnetic field and discard the supernatant. Wash the magnetic balls twice with 1 mL of 70% ethanol solution, magnetically separate, discard the supernatant, and place the magnetic balls adsorbing DNA at room temperature to volatilize the ethanol.

[0063] Add 0.4mL of 2mol / L NaCl solution to the centrifuge tube containing the magnetic ball, shake well, place at room temperature for 30 minutes until D...

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Abstract

The invention discloses a preparation method of core-shell magnetic conductive polymer microspheres. Ferrite (Fe3O4, gamma-Fe2O3, MFe2O4; M = Fe, Co, Ni, Mn and the like) nano or micro spherical or near-spherical particles are used as cores; firstly, surface hydroxyl protonation is performed on the magnetic cores to fully combine the magnetic cores before compositing with monomer molecules by theelectrostatic interaction and hydrogen-bond action between the protonated hydroxy and monomer aniline, pyrrole or derivatives thereof; and in the oxidation polymerization process, the monomer molecules are controlled to form cores and grow on the surfaces of the magnetic cores, thereby preparing the core-shell magnetic conductive polymer microspheres with high magnetic responsibility and good dispersity. By the characteristics that the amido-containing magnetic polymer microspheres are positively charged in a high-pH solution while the DNA molecules in a tissue or cell lysis solution are negatively charged, the prepared magnetic composite microspheres are applied to DNA separation and purification assisted by an external magnetic field.

Description

Technical field [0001] The invention relates to a method for preparing core-shell magnetic conductive polymer microspheres and its application in DNA separation and purification, and belongs to the technical field of magnetic composite materials and biological separation materials preparation. Background technique [0002] The separation and purification of DNA has a very important position in the fields of genetic engineering, biology, medicine and genetics. DNA separation and purification technologies can be generally divided into two categories: no external magnetic field assistance and external magnetic field assistance, namely non-magnetic separation and magnetic separation. There are many methods of non-magnetic separation, which are described in many documents and patents (N. Ramashupuramanyan Chinese Patent: DNA Purification Method and Purified DNA. CN 1390229A, January 8, 2003), Such as: phenol-chloroform-isoamyl alcohol method, cesium chloride-ethidium bromide density ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08G73/02C08G73/06C08K9/02C08K3/22C12N15/10
Inventor 盖利刚马万勇姜海辉韩小云
Owner QILU UNIV OF TECH
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