Magnetically responsive DNA separation medium and preparation and use methods thereof

A separation medium, magnetic responsive technology, applied in the medium of Al3+ chelation separation DNA, magnetic responsive DNA extraction and separation medium, can solve the problems of low selectivity, take a long time, etc., achieve good selectivity, Shorten the extraction and separation time, the effect of small particle size

Pending Publication Date: 2021-11-02
BEIJING UNIV OF CHEM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The basic principle of separating DNA from the two is the adsorption of silicon-based media on DNA, but the selectivity of this type of media for DNA adsorption is not strong. High concentrations of ethanol, isopropanol and other organic reagents are used in the extraction process to remove or volatilize these Organic reagents take longer

Method used

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  • Magnetically responsive DNA separation medium and preparation and use methods thereof
  • Magnetically responsive DNA separation medium and preparation and use methods thereof
  • Magnetically responsive DNA separation medium and preparation and use methods thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0020] (1) Weigh 4g of FeCl 3 ·6H 2 O, add 80 mL of ethylene glycol, stir magnetically until it is completely dissolved to form a clear solution, then add 12 g of anhydrous sodium acetate and 3 g of polyethylene glycol-4000, and magnetically stir to dissolve all of it to obtain a mixed solution. Then, the mixed liquid was poured into a stainless steel reaction kettle with a polytetrafluoroethylene substrate and sealed, and the reaction was carried out at 190° C. for 12 h. After the reaction, it was naturally cooled to room temperature, and the obtained product was magnetically separated, washed three times with deionized water and absolute ethanol, and dried under vacuum at 60 °C for 6 h to obtain Fe. 3 O 4 magnetic particles.

[0021] (2) Take 50mL of NaOH solution of 1.0mol / L, add 25mL of 1,4-dioxane and 5mL of epichlorohydrin successively, mix well to obtain mixed solution A, then add 2g of mixed solution A according to the present embodiment The Fe obtained by the proc...

Embodiment 2

[0027] (1) Take 0.5g Fe of modified IDA prepared by the process method of step (1) and step (2) in Example 1 3 O 4 Magnetic particles, added to 60 mL of 1.0 mol / L Al(NO 3 ) 3 The solution was mixed well, placed in a shaker at 50 °C for 2 hours, and after the reaction was completed, the solid material separated under the action of a magnetic field was washed three times with deionized water, and dried in vacuum at 60 °C for 6 hours to obtain bonded Al 3+ Fe 3 O 4 Magnetic particles are the DNA separation medium Fe 3 O 4 @Al 3+ , ICP-AES detects Fe 3 O 4 @Al 3+ Medium Al 3+ The content was 0.096 mmol / g.

[0028] (2) Take 700 μL of the cell lysis serum containing plasmid DNA prepared by the method of step (5) of Example 1, and add 10 mg of Fe prepared in step (1) of this example. 3 O 4 @Al 3+ , Mix well, carry out magnetic field separation after adsorption reaction at room temperature for 15 minutes, add 400 μL of washing solution (10 mmol / L Tris-HCl, pH=8.0) to the...

Embodiment 3

[0031] (1) Take 1 g of Fe of modified IDA prepared by the process method of step (1) and step (2) in Example 1 3 O 4 Magnetic particles, added to 40mL of 0.5mol / L Al(NO 3 ) 3 The solution was placed in a shaker at 20 °C for 22 hours of shaking reaction. After the reaction was completed, the solid material was obtained by magnetic field separation and washed three times with deionized water, and was vacuum dried at 60 °C for 6 hours to obtain bonded Al. 3+ Fe 3 O 4 Magnetic particles are the DNA separation medium Fe 3 O 4 @Al 3+ . ICP-AES detection of Al 3+ The content is 0.14mmol / g Fe 3 O 4 @Al 3+ .

[0032] (2) Take 700 μL of the plasmid DNA-containing cell lysate obtained by the process method in step (5) of Example 1, and add 5 mg of Fe prepared in step (1) of this example. 3 O 4 @Al 3+ , Mix well, carry out magnetic field separation after adsorption reaction at room temperature for 15 min, add 400 μL of washing solution (10 mmol / LTris-HCl, pH=8.0) to the obt...

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Abstract

The invention discloses a magnetically responsive DNA separation medium and a preparation and use method thereof, and belongs to the technical field of biochemical separation medium preparation and application. The magnetically responsive DNA separation medium (Fe3O4@Al<3+>) is structurally characterized in that aluminum ions (Al<3+>) are bonded on Fe3O4 magnetic particles which are activated by epichlorohydrin and modified with iminodiacetic acid. The Fe3O4@Al<3+> has monodispersity, large specific surface area and simple preparation process, and selective separation of the Fe3O4@Al<3+> on DNA is realized based on metal chelation of Al<3+> on DNA; the DNA extraction and separation process is simple, convenient and rapid, and high-flux rapid extraction of DNA molecules is easy to realize.

Description

technical field [0001] The invention belongs to a magnetically responsive DNA extraction and separation medium and a preparation and use method thereof, and belongs to the technical field of extraction and separation of biochemical substances, in particular to a magnetically responsive Al 3+ A medium for DNA separation by chelation (Fe 3 O 4 @Al 3 + ) and methods for their preparation and use. Background technique [0002] DNA is widely present as the main genetic material in living organisms. Extracting and isolating DNA is the basis and premise of research in life sciences and related fields. There are many methods for DNA extraction and separation, but limited by the extraction efficiency and cost, the commonly used methods currently mainly include silica membrane spin column method and magnetic bead method. The silica membrane spin column method mostly uses a silica-based column to adsorb DNA, and combines centrifugation to extract and separate the DNA. Most of the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1013
Inventor 刘长霞陈驰杨文胜
Owner BEIJING UNIV OF CHEM TECH
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