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Polymer/gold nano particle composite medium for use in capillary electrophoresis DNA sequencing and method for preparing same

A DNA sequencing and gold nanoparticle technology, applied in the field of polymer/gold nanoparticle composite media and its preparation, can solve the problems of inability to sequence DNA, low viscosity, etc., achieve easy injection and flushing out of capillaries, and reduce polymer degradation possibility, viscosity reduction effect

Inactive Publication Date: 2010-06-23
UNIV OF SCI & TECH OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But also due to the extremely low viscosity, this medium cannot sort DNA

Method used

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  • Polymer/gold nano particle composite medium for use in capillary electrophoresis DNA sequencing and method for preparing same
  • Polymer/gold nano particle composite medium for use in capillary electrophoresis DNA sequencing and method for preparing same
  • Polymer/gold nano particle composite medium for use in capillary electrophoresis DNA sequencing and method for preparing same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] (1) Preparation of LPA / PDMA quasi-interpenetrating polymer network

[0042] Add Span 80 (2.47g) and refined kerosene (50mL) into a 250mL clean and dry four-neck round bottom flask, and install a mechanical stirrer, reflux condenser, nitrogen inlet / outlet tube and constant pressure funnel on the flask . Acrylamide (AM, 20g) and H 2 A solution of O (30 g) was added dropwise to the mixture. Under the condition of a mechanical stirring speed of 500 rpm, ultra-high purity nitrogen (UHP, 99.99%) was continuously introduced into the system for 1 h, and then 40 μL of APS (0.1 g / mL) aqueous solution and 4.5 μL of TEMED were added, and reacted at 22 ° C for 24 h. After the polymerization, the emulsion was precipitated in excess acetone, filtered, vacuum-dried, dissolved in water, precipitated with acetone, and vacuum-dried three times to obtain purified linear polyacrylamide (LPA). The viscosity-average molecular weight of LPA measured by Ubbelohde viscometer is 3.0×10 6 Da. ...

Embodiment 2

[0053] (1) Preparation of LPA / PDMA quasi-interpenetrating network

[0054] Method is the same as embodiment one.

[0055] (2) Preparation of 40nm Au particles

[0056] Method is the same as embodiment one.

[0057] Preparation and preparation of sieving media quasi-IPN / GNPs-2 with the same particle size and different content of Au

[0058] (3) Preparation and formulation of sieving medium quasi-IPN / GNPs-1

[0059] 2.0 mL of the above-mentioned Au sol was added to 20 mL of the above-mentioned 1% (w / v) quasi-IPN aqueous solution, respectively. After complete mixing, the solution was precipitated with excess acetone, filtered and dried in vacuo.

[0060] When performing DNA sequencing experiments, the medium was dissolved with 1×TTE buffer solution and diluted to 2.5% (the concentration unit of the polymer solution is w / v).

[0061] The implementation conditions of capillary electrophoresis are the same as in Example 1.

[0062] 2.5% quasi-IPN / GNPs-2 is used as the DNA sequ...

Embodiment 3

[0064](1) Preparation of LPA / PDMA quasi-interpenetrating network

[0065] Method is the same as embodiment one.

[0066] (2) Preparation of 20nm Au particles

[0067] All required glassware was cleaned with aqua regia, rinsed with deionized water and dried. Add 50 mL of 0.01% (wt.) HAuCl to a round bottom flask equipped with a condenser 4 Aqueous solution, the solution was heated to boiling under vigorous stirring conditions. Quickly add 0.85 mL of 1% (wt.) sodium citrate aqueous solution to the above boiling solution, the color of the solution changes from light yellow to blue quickly, and then to purple red. After boiling for 10 minutes, remove the heat source. The sol was then stirred for 15 min and then cooled to room temperature. TEM results show that the average diameter of Au is 20nm.

[0068] (3) Preparation and formulation of quasi-IPN / GNPs-3 sieving medium with Au particle size of 20nm

[0069] 1.0 mL of the above-mentioned Au sol was added to 20 mL of the abov...

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Abstract

The invention relates to polymers / gold nanoparticles complex medium for DNA sequencing with capillary electrophoresis and its preparation method, and belongs to the biotechnological field of DNA sequencing and separation. The concentration of the buffer solution of polymers / gold nanoparticles complex medium used for DNA sequencing is 2.0%-3.0%. The invention comprises the steps of: (1) synthesis of quasi-interpenetrating polymer networks with self-spreading ability, (2) preparation of Au nanoparticles, and (3) synthesis of complex medium of quasi-interpenetrating polymer networks / gold nanoparticles. The complex medium with reduced viscosity at proper concentration is easy to be pumped into and run out of the capillaries, and can prevent the production of electroosmotic flow because of theexistence of complex with self spreading ability, which saves the step of spreading inner wall of the capillary, simplifies the preparation process of column, reduces the application cost of capillary, elongates the service time of capillary. The invention is mainly used for separating DNA with capillary electrophoresis and DNA sequencing.

Description

technical field [0001] The invention relates to a polymer / gold nanoparticle composite medium for capillary electrophoresis DNA sequencing and a preparation method thereof, belonging to the technical field of DNA sequencing and separation biotechnology. Background technique [0002] Capillary electrophoresis is a well-established method for DNA analysis. For capillary electrophoresis, since the separation medium determines the migration behavior and separation degree of DNA fragments, it is an important factor for DNA separation. Currently, synthetic polymers are the most widely used separation medium for DNA separation. For an effective For separation media, it should have high screening capacity, low viscosity and self-coating function at the same time. [0003] Synthetic polymers commonly used to separate DNA mainly include hydrophilic homopolymers (Heller C., Electrophoresis, 2001, 22: 629-643). For these homopolymers, most of them need to coat the inner wall of the capi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C08L33/24C08K3/08
Inventor 王延梅周丹
Owner UNIV OF SCI & TECH OF CHINA
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