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Polymer for capillary electrophoresis and preparation method thereof

A technology of capillary electrophoresis and polymers, which is applied in separation methods, chemical instruments and methods, and material analysis by electromagnetic means, can solve the problems that the stability of gel sieving properties and other physical properties cannot be guaranteed, and prevent analysis The effect of sticking to the wall

Active Publication Date: 2014-11-05
INST OF FORENSIC SCI OF MIN OF PUBLIC SECURITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If the molecular weight of the polymer cannot be effectively controlled, the stability of the sieving properties of the gel and the stability of other physical properties cannot be guaranteed.

Method used

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  • Polymer for capillary electrophoresis and preparation method thereof
  • Polymer for capillary electrophoresis and preparation method thereof
  • Polymer for capillary electrophoresis and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1, the preparation of gel

[0029] 1. Preparation of sieving medium

[0030] 1. Polymerization reaction

[0031] Add 30 g of N,N-dimethylacrylamide monomer (DMA), 60 mL of dioxane, 0.15 g of azobisisobutyronitrile (AIBN) and a stirring bar in a round bottom flask, cover with a rubber stopper, and stir uniform.

[0032] Argon was blown for 1 hour to remove oxygen, and then reacted at 55° C. for 15 minutes. The system became viscous to stop the reaction, and a polymerized solution was obtained.

[0033] 2. Post-processing (purification and drying):

[0034] 1) Purification:

[0035] Polymerized solution is suitably diluted and then precipitated and separated three times with n-hexane (dissolved in dichloromethane). The specific steps are:

[0036] The polymer in the polymerized solution is precipitated with n-hexane, and the supernatant is removed to obtain the first precipitated product; the first precipitated product is dissolved in methylene chloride, a...

Embodiment 2

[0055] Embodiment 2, gel application on AB 310 type genetic analyzer

[0056] 1. Electrophoresis conditions on the AB 310 genetic analyzer;

[0057] Electrophoresis voltage is 15kV;

[0058] The capillary length is 50cm;

[0059] The electrophoresis temperature was 60°C.

[0060] 2. Replace with newly prepared Running buffer (A.C.E, AMRESCO company, diluted 10 times); internal standard (2 μL internal standard NH 500 (purchased from Suzhou Xinhai Biotechnology Co., Ltd.) + 23 μL deionized formamide).

[0061] 3. Draw up the glue (about 0.6mL) with a syringe, install it to a fixed position, open the buffer valve, and remove the air bubbles in the tube.

[0062] 4. Manual glue removal.

[0063] 5. Detection by capillary electrophoresis

[0064] Test results such as figure 2 As shown, the peak shape of the internal standard is sharp and the resolution is good, indicating that the prepared gel can form a good separation of the 17 DNA fragments of NH-500.

Embodiment 3

[0065] Example 3, Typer TM Detection of 500 molecular weight internal standard

[0066] The gel was applied on an AB 3130xl genetic analyzer.

[0067] 1. Electrophoresis conditions for the separation of DNA fragments on the AB 3130 genetic analyzer

[0068] Electrophoresis voltage is 15kV;

[0069] Injection time is 10s

[0070] Capillary length is 36cm;

[0071] The electrophoresis temperature was 60°C.

[0072] 2. Replace the newly prepared Running buffer (A.C.E, AMRESCO company, diluted 10 times) and internal standard (1μL Typer TM 500 internal standard (purchased from the Physical Evidence Identification Center of the Ministry of Public Security)) + 20 μL deionized formamide).

[0073] 3. Put the gel prepared by the present invention into a glue bottle, place the glue injection tube in the glue bottle, and fix it.

[0074] 4. Start the automatic rubber changing system.

[0075] 5. Perform capillary electrophoresis detection according to the operating instructions ...

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Abstract

The invention discloses a polymer for capillary electrophoresis and a preparation method thereof. The method comprises the following steps of: mixing polymer monomer, a solvent and an initiator, carrying out polymerization reaction under the anaerobic environment and obtaining a sieving medium; the time of the polymerization reaction is 10-35 minutes, and the temperature of the polymerization reaction is 45-65 DEG C; and the mixing ratio of the polymer monomer, the solvent and the initiator is (25-35)g:(50-70)ml:(0.1-0.2)g. According to the method disclosed by the invention, enough amount of polymer is absorbed on a silicon surface to establish a high-viscosity area on the silicon surface, the movement of double electric layers under the action of the electric field can be hindered, and analytes are prevented from being attached with the wall. In the invention, the formed polymer with constant viscosity is filled in a capillary tube, the sieving of the substances to be analyzed can be effectively realized, and the generation of an electroosmosis site can be avoided to a great extent.

Description

technical field [0001] The invention relates to a polymer for capillary electrophoresis and a preparation method thereof. Background technique [0002] Capillary electrophoresis is a kind of liquid phase micro-separation analysis technology that uses capillary as the separation channel, uses high-voltage direct current electric field as the driving force, and achieves the purpose of separation based on the different moving speeds of samples under the electric field. It has high separation efficiency, short analysis time, and low sample consumption. Less advantages. Capillary electrophoresis includes capillary gel electrophoresis (CGE) and capillary gel-free sizing electrophoresis (NGS). The former uses a cross-linked polymer gel, while the latter uses a hydrophilic linear polymer solution (also called a non-cross-linked polymer solution) as a sieving medium, so it is called gel-free sieving electrophoresis. Gel-free sieving electrophoresis avoids the complicated gel prepar...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08F120/56C08J3/075G01N27/447B01D57/02
Inventor 赵兴春叶健毕万里
Owner INST OF FORENSIC SCI OF MIN OF PUBLIC SECURITY