Method for preparing double-epoxy modified biochip substrate

A technology of bis-epoxy and biochips, applied in biochemical equipment and methods, measurement/inspection of microorganisms, instruments, etc., can solve the problems of not improving the binding force and detection signal strength, and achieve simple preparation methods Easy to operate, regular spotting, and strong binding effect

Inactive Publication Date: 2013-01-02
DONGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the secondary modification of amino-modified biochip substrates with aldehyde groups has been optimized to a certain extent, but the binding force and detection signal intensity have not been improved very well.

Method used

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  • Method for preparing double-epoxy modified biochip substrate
  • Method for preparing double-epoxy modified biochip substrate
  • Method for preparing double-epoxy modified biochip substrate

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] (1) Measure 115.7ml of 99.8% absolute ethanol, 7.5ml of silane coupling agent, and 1.8ml of glacial acetic acid into a three-necked flask, and stir magnetically for 60 minutes at room temperature.

[0025] (2) Put the glass piece into a dyeing jar filled with ultrapure water and ultrasonically for 10 minutes, wash it twice with ultrapure water, and then soak it in concentrated sulfuric acid, hydrogen peroxide and ultrapure water (volume ratio is 4:1:20 ) in a mixed solution of 130°C for 30 minutes, cooled, rinsed 3 times with ultrapure water; then immersed in a mixed solution of concentrated hydrochloric acid and ethanol (1:1 by volume) for 24 hours, Wash with ultrapure water 5 times, dry at 110°C for 30 minutes, and cool to room temperature.

[0026] (3) Put the surface-treated glass sheet into the aminosilane coupling agent solution, then put it into a constant temperature and humidity box, control the relative humidity at 50%, set the temperature at 30°C, and place i...

Embodiment 2

[0029] (1) Measure 122.5ml of 99.8% absolute ethanol, 1.9ml of silane coupling agent, and 0.5ml of glacial acetic acid into a three-necked flask, and stir magnetically at room temperature for 90 minutes.

[0030] (2) Put the glass piece into a dyeing jar filled with ultrapure water and ultrasonically for 5 minutes, wash it twice with ultrapure water, and then soak it in concentrated sulfuric acid, hydrogen peroxide and ultrapure water (volume ratio is 4:1:20 ) in a mixed solution of 80°C for 60 minutes, cooled, rinsed 3 times with ultrapure water; then soaked in a mixed solution of concentrated hydrochloric acid and ethanol (1:1 by volume) for 3 hours, Wash with ultrapure water three times, dry at 140°C for 15 minutes, and cool to room temperature.

[0031] (3) Put the surface-treated glass sheet into the aminosilane coupling agent solution, then put it into a constant temperature and humidity box, control the relative humidity at 30%, set the temperature at 25°C, and place it...

Embodiment 3

[0034] Detection of Immobilization Ability of Diepoxide-Modified Biochip Substrate

[0035] The biochip substrate modified by the double epoxy group used in this example was prepared according to the method described in Example 1, and this example was obtained by Arrayit 2. The spotting instrument performs spotting test on the epoxy substrate, and the schematic diagram of spotting matrix design is as follows: figure 2 shown, and then through the Axon 4000B: Cy3 PMT600, Power100 scanner detects its fluorescence signal intensity to reflect the immobilization ability of the bis-epoxy modified biochip substrate, the specific operation is as follows:

[0036] (1) Prepare samples for spotting

[0037] a. At room temperature, add DNA probes to 50% DMSO / 50% H 2 O buffer. Add antibody probes to 20% glycerol / 80% H 2 O buffer;

[0038] b. Transfer the sample to a 96-well plate or a 384-well plate according to the schematic diagram of the matrix design, tap the microplate to make...

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Abstract

The invention relates to a method for preparing a double-epoxy modified biochip substrate, which comprises processes of surface treatment, preparation of silane coupling agent solution, amino modification, preparation of double-epoxy reagent solution and double-epoxy modification. One of two epoxy groups in a double-epoxy reagent is reacted with an amino on the substrate and opened, and the otherepoxy group is exposed on the surface of the substrate. The double-epoxy modified biochip substrate well solves the problems of small acting force of amino substrate contacts and non-uniform point samples, promotes the signal intensity of a chip to a great extent compared with an aldehyde group modified biochip substrate, and can be widely applied to various biochips.

Description

technical field [0001] The invention belongs to the field of preparation of biochip substrates, in particular to a method for preparing biochip substrates modified by double epoxy groups. Background technique [0002] Biochip technology uses microfabrication technology to process microstructures for the preparation, reaction and detection of biological samples on glass, plastic or other matrix materials. Accurate, fast, and large-information detection and analysis of genes, ligands, antigens and other biologically active substances greatly save time and cost. [0003] The immobilization of oligonucleotides, gene fragments or protein antigens and antibodies on the surface of the substrate is a prerequisite for the production of biochips. One of the key technologies of biochips. The surface of the biochip substrate should have active functional groups that can conduct chemical reactions and couple with biomolecules, and the substrate itself should have sufficient stability a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68G01N33/552
Inventor 王宏志崔博马董云张青红李耀刚
Owner DONGHUA UNIV
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