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Precipitation-oxidation method for preparing glucan ferroferric oxide magnetic nano particle and application thereof

A technology of magnetic nanoparticles and ferric oxide, which is applied in the fields of nanomaterials and bioengineering, can solve problems such as difficult control and complicated steps, and achieve the effects of simple operation, improved sensitivity, and simplified conditions.

Inactive Publication Date: 2010-02-03
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For the extraction of genetically modified soybean grain DNA, some traditional DNA separation and purification methods such as the CTAB method are generally used, which not only need to contact a variety of organic solvents and toxic reagents, but also the steps are complicated and difficult to control

Method used

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  • Precipitation-oxidation method for preparing glucan ferroferric oxide magnetic nano particle and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1. The preparation of dextran ferroferric oxide magnetic nanoparticles, the specific steps are as follows:

[0031] (1) 5g of Dextran T-40 was completely dissolved in 10mL of deionized water under nitrogen protection and mechanically stirred at 200rpm.

[0032] (2) 4mmol FeCl 2 4H 2 O was dissolved in 10 mL of deionized water under nitrogen protection, slowly dropped into the dextran solution and continued to stir.

[0033] (3) Heat up the water bath to 60°C, stir at 350rpm for 5min, slowly add 10mL of 25%-28% ammonia water dropwise, the reaction solution turns blue, remove the nitrogen protection, and expose the reaction system to air.

[0034] (4) After 20 minutes, the reaction solution gradually changed from blue to black, and finally a black colloidal solution was obtained. Stop the reaction and cool for 30 minutes.

[0035] (5) Adjust the pH of the black colloid to 7.0 with glacial acetic acid, centrifuge at 8000 rpm for 10 min, discard the precipitat...

Embodiment 2

[0036] Example 2. The extraction and purification of transgenic soybean DNA, the steps are as follows:

[0037] (1) Wash 3 transgenic soybean seeds, soak them in pure water for 24 hours, peel them manually, add a small amount of quartz sand, grind them into a paste in an ice bath, and transfer them to a 10mL centrifuge tube.

[0038](2) Quickly add 65°C preheated extract solution (0.1M Tris-HCl pH8.0, 0.02M EDTA pH8.0, 1.5M NaCl, 2% PVP 40, 2% CTAB, 3% β-mercaptoethanol) to 9mL, shake up and down, 65 ℃ water bath for 1h, invert three times every 5min, centrifuge at 10000rpm for 10min, take the supernatant.

[0039] (3) Add an equal volume of chloroform-isoamyl alcohol (24:1, V / V) to the supernatant and slowly invert it up and down 30 times, centrifuge at 10,000 rpm for 10 min, take 1 mL of the supernatant and add it to a 1.5 mL centrifuge tube.

[0040] (4) Add 10 μL of 10 mg / mL dextran ferroferric oxide magnetic nanoparticles, magnetically separate the precipitate, wash 3 ti...

Embodiment 3

[0041] The purification of embodiment 3.PCR product, steps are as follows:

[0042] Magnetic nanoparticles with a solid content of 10 mg / mL were ultrasonically dispersed for 5 min, 15 μL of magnetic nanoparticle dispersion was added to a PCR reaction tube, 20 μL of hybridization solution (10% PEG-8000, 0.2M NaCl) was added, incubated at 37°C for 5 min, and magnetically separated , discard the supernatant, wash the magnetic particles with 20 μL of ethanol with a mass concentration of 70% for 2 to 3 times and air-dry for 2 minutes, then elute the DNA with 20 μL of Tris-EDTA eluent, and measure the OD by ultraviolet light 260 / OD 280 Then take 2 μL for agarose gel electrophoresis and determine the DNA sequence.

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Abstract

The invention relates to a precipitation-oxidation method for preparing aglucan ferroferric oxide magnetic nano particle and an application thereof, belonging to the technical fields of nano materialsand bioengineering. The invention comprises a method for preparing a glucan ferroferric oxide magnetic nano particle used for extraction and purification of DNA and the purification of PCR products in the process of transgenic soy detection. The invention provides a simple, convenient and easy precipitation-oxidation one-step method for preparing the glucan ferroferric oxide magnetic nano particle, simplifies the reaction condition, obtains spherical particle with larger particle diameter, high magnetic response and favorable water solubility; the prepared magnetic nano particle is used for the efficient extraction and purification of the DNA and the purification of PCR products in the process of transgenic soy detection, greatly improves the sensitivity of detection and is simpler and more convenient to operate.

Description

technical field [0001] A method for preparing superparamagnetic dextran ferroferric oxide magnetic nanoparticles and its application in transgene detection belongs 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] Glucan, also known as dextran, is a homopolysaccharide formed by the polymerization of several glucose molecules, mainly connected by D-glucopyranose with α-(1,6) glycosidic bonds, itself and its degradation products in the body ( CO 2 、H 2 O) It is non-toxic and harmless to the human body. Dextran is easily soluble in water and can replace a part of ...

Claims

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Application Information

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IPC IPC(8): H01F1/10H01F1/34C12N15/10C01G49/08C08L5/02C08K3/22
Inventor 胥传来刘微波陈伟赵媛彭池方
Owner JIANGNAN UNIV
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