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Preparation of polyacrylic acid coated ferroferric oxide magnetic nano-particles and application thereof

A technology of magnetic nanoparticles and ferric oxide, applied in the fields of nanomaterials and bioengineering, can solve the problems of complicated steps and difficult control, and achieve the effect of simple operation and improved extraction rate.

Inactive Publication Date: 2009-12-30
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Traditional DNA separation and purification methods not only need to contact with a variety of organic solvents and toxic reagents, but also the steps are complicated and difficult to control

Method used

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  • Preparation of polyacrylic acid coated ferroferric oxide magnetic nano-particles and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0023] The preparation of embodiment 1PAA ferroferric oxide magnetic nanoparticles

[0024] It is synthesized by pyrolysis method, and its chemical reaction is:

[0025] FeCl 3 →Fe(OH) 3 →Fe(OH) 2 → Fe 3 o 4

[0026] Proceed as follows:

[0027] (1) Weigh 4g NaOH (100mmol) and dissolve it in 20mL diethylene glycol (DEG), heat in an oil bath at 120°C for 1h under the protection of nitrogen, stir to completely dissolve NaOH, and obtain a light yellow colloidal solution, which is sealed and stored in a water bath at 70°C .

[0028] (2) Weigh 720mg PAA (10mmol) and 648.84mg FeCl 3 (4mmol) powder, dissolved in 20mL DEG, heated and refluxed in an oil bath at 220°C for 2h under the protection of nitrogen, added the DEG liquid of 15mL NaOH that had been prepared and stored, and continued to heat and reflux for 2h to obtain a black viscous colloidal liquid, and terminate the reaction. Let stand to cool.

[0029] (3) Take 2 mL of the reaction solution in a centrifuge tube, add...

Embodiment 2

[0032] Example 2 Extraction and Purification of Cellular DNA

[0033] Proceed as follows:

[0034] (1) Centrifuge 2 mL of cells at 1000 rpm for 5 min, discard the supernatant, add 5 mL of normal saline, and centrifuge twice;

[0035] (2) Add 5 μL cell lysate (3M NaCl, 5M Urea, 40g / L TritonX-100, 10mM EDTA, 25mM Tris-HCl pH6.5) to the centrifuged cells and mix well, add 10μL proteinase K (25μg / μL) and 5 μL RNase (40 μg / μL), incubate at 37°C for 30 minutes, add 10 μL magnetic particles (18 mg / mL) and let stand for 2 minutes, add 20 μL of isopropanol and let stand for 2 minutes, magnetically separate, add 20 μL of 70% ethanol, magnetically separate, discard To the supernatant, add 20 μL of Tris-EDTA buffer, incubate at 37°C for 5 min, magnetically separate, and take the supernatant, which is the DNA eluent.

[0036] (3) OD measured by ultraviolet light 260 / OD 280 The ratio, and take 2μL agarose gel electrophoresis.

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Abstract

The invention discloses preparation of polyacrylic acid (PAA) coated ferroferric oxide magnetic nano-particles and application thereof, and belongs to the technical fields of nano materials and bioengineering. The invention comprises a method for preparing the PAA ferroferric oxide magnetic nano-particles and application of the PAA ferroferric oxide magnetic nano-particles in cell DNA extraction and purification. The invention provides a method for preparing the PAA ferroferric oxide magnetic nano-particles by a high temperature decomposition method; and the prepared magnetic nano-particles are used for cell DNA high-efficiency extraction and purification, the extraction ratio is greatly improved, and the operation is more simple and convenient. The separation efficiency and the DNA yield of the DNA purification method based on the magnetic nano-particles are not lower than those of the conventional method.

Description

technical field [0001] The invention discloses a preparation method and application of polyacrylic acid (PAA)-coated ferroferric oxide magnetic nanoparticles, belonging to the technical field of nanomaterials and bioengineering. Background technique [0002] Ferroferric oxide magnetic nanoparticles have low toxicity and magnetic responsiveness. When the particle size is small enough, they have superparamagnetism, that is, they have strong magnetism in an external magnetic field. When the magnetic field is withdrawn, the magnetism disappears quickly and the residual magnetism is zero. , will not be permanently magnetized. [0003] The ferroferric oxide magnetic nanoparticles coated with PAA can make it not only magnetic and water-soluble, but also have surface active groups, which can be further coupled with various biomolecules such as cells, enzymes, proteins, antibodies and nucleic acids. Under the action of an external magnetic field, the magnetic nanoparticles can be ea...

Claims

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Application Information

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IPC IPC(8): B01J13/02B01J20/26B01J20/28B01J20/30C07H1/06C07H21/04H01F1/11
Inventor 胥传来刘微波陈伟赵媛彭池方匡华
Owner JIANGNAN UNIV
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