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A gel-free sieving medium for capillary electrophoresis and its preparation method

A technology of capillary electrophoresis and sieving, which is applied in biochemical equipment and methods, DNA preparation, microbial measurement/inspection, etc. It can solve the problems of poor stability and low resolution, achieve high hydrophilicity, reduce adsorption, Ease of injection and replacement of media

Active Publication Date: 2018-02-02
THE FIRST RES INST OF MIN OF PUBLIC SECURITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the patent still has the problems of low resolution and poor stability

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  • A gel-free sieving medium for capillary electrophoresis and its preparation method
  • A gel-free sieving medium for capillary electrophoresis and its preparation method
  • A gel-free sieving medium for capillary electrophoresis and its preparation method

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Embodiment 1

[0030] A kind of glue-free sieving medium for capillary electrophoresis provided by the present invention comprises polymer monomer, solvent, chain transfer agent, initiator and catalyst, described polymer monomer is acrylamide, and described solvent is Deionized water, the chain transfer agent is Virahol, the initiator is ammonium persulfate (APS), the catalyst is N,N,N,N-tetramethyldiethylamine (TEMED), the The ratio of acrylamide, deionized water, isopropanol, ammonium persulfate (APS) and N,N,N,N-tetramethyldiethylamine (TEMED) is (20-30)g: (180-270 )ml: (5.24-7.86)ml: (1.0-2.0)ml: (1.0-2.0)ml, to obtain a glue-free sieving medium, the acrylamide, deionized water, isopropanol, ammonium persulfate (APS) The ratio of N,N,N,N-tetramethyldiethylamine (TEMED) is 25g: 222ml: 6.55ml: 1.25ml: 1.25ml, preferably. Described glue-free sieving medium is polyacrylamide (LPA), and structural formula is as follows:

[0031]

[0032] The present invention also provides a preparation ...

Embodiment 2

[0038] In this example, the reagents used are mainly from Sigma-Aldrich Company (USA). The specific operation is prepared according to the following steps: add 222ml of deionized water and 6.55ml of isopropanol (analytical grade) to a round bottom flask, and feed high-purity nitrogen (99.99%) at 35°C to deoxygenate the solution for 10 minutes. Then add 25g of acrylamide (AM), 1.25ml of 10% (v / v) N,N,N,N-tetramethyldiethylamine (TEMED) and 1.25ml of 10% (w / v) persulfuric acid Ammonium (APS), magnetically stirred at 500rpm, after reacting for 90min, the solution was honey viscous. A dialysis bag (Spectra / por#2, Spectrum Corporation, USA) with a molecular weight cut-off of 12-14 KDa was selected for dialysis for three days. After dialysis, the polymer solution was freeze-dried at -60°C for 72 hours to obtain a white solid, which was the sieving medium, and the reaction yield was about 80%.

Embodiment 3

[0039] The characterization of embodiment 3 sieving media

[0040] 1 1 H-NMR spectrometer: with D 2 O was used as a solvent to characterize the polymer (white solid) as figure 1 shown. from figure 1 It can be seen that the structure of the polymer (polyacrylamide) is correct, and the purity is high, and it does not contain impurities such as monomers and solvents.

[0041] 2 Gel Permeation Chromatography (GPC): The polymer was subjected to Waters Breeze TM 2 HPLC system test and analysis, the column is Waters Ultrahydrogel Linear Column, the pump is Waters 1515, the PDA detector, the mobile phase is water, the test results are shown in Table 1.

[0042] Table 1 Gel Permeation Chromatography (GPC) Test Results

[0043]

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Abstract

The invention relates to the technical field of capillary electrophoreses, in particular to a non-gel sieving medium for capillary electrophoresis and a preparation method thereof. The non-gel sieving medium for capillary electrophoresis comprises a polymer monomer which is acrylamide, solvent which is deionized water, a chain transfer agent which is isopropanol, an initiator which is ammonium persulfate and a catalyst which is an N,N,N,N-tetramethylethylenediamine mixture. The preparation method of the non-gel sieving medium for capillary electrophoresis comprises the steps that the polymer monomer, the solvent, the chain transfer agent, the initiator and the catalyst are mixed, and after dialysis and freeze drying are conducted, the mixture is swelled in a sol buffer solution, wherein the sieving medium is polyacrylamide. The prepared sieving medium has good sieving performance, dissolving capacity and high hydrophilicity, the adsorptive action between DNA and the capillary wall is effectively reduced, the high molecular weight and low viscosity are achieved, medium injection and replacement are convenient, automation of DNA analysis and separation is facilitated, and the requirements of legal medical experts on STR analysis and detection can be met.

Description

technical field [0001] The invention relates to the technical field of sieving media for capillary electrophoresis, in particular to a gel-free sieving medium for capillary electrophoresis and a preparation method thereof. Background technique [0002] In the prior art, capillary electrophoresis (capillary electrophoresis, CE), also called high-performance capillary electrophoresis (HPCE), is one of the fastest-growing analytical methods in recent years. In 1981, Jorgenson and Lukacs first proposed to separate by applying high voltage in a capillary column with an inner diameter of 75 μm, and created modern capillary electrophoresis. In 1984, Terabe et al established micellar capillary electrokinetic chromatography. In 1987, Hjerten established capillary isoelectric focusing, and Cohen and Karger proposed capillary gel electrophoresis. From 1988 to 1989, the first batch of commercial capillary electrophoresis instruments appeared. In just a few years, because capillary el...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08F120/56C08F2/38C08F4/30C12N15/10C12Q1/68
Inventor 俞丽娟聂燕钗姜伯玮
Owner THE FIRST RES INST OF MIN OF PUBLIC SECURITY
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