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Chip preparation method, DNA or protein immobilization method and chip

A DNA chip and protein chip technology, applied in the field of chip preparation, can solve the problems of poor repeatability of DNA chip detection, high false positive or false negative, wrong sequence information, etc., to achieve high controllability, improve quality and efficiency, and repeatability Good results

Inactive Publication Date: 2019-04-12
GENEMIND BIOSCIENCES CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] DNA chips with unreliable DNA immobilization are prone to the following defects: 1) low detection rate; 2) high false positive or false negative; 3) give wrong sequence information; 4) poor repeatability of DNA chip detection
However, as an active chemical reaction, covalent linkage faces many problems in terms of controllability, speed and repeatability of the reaction.

Method used

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  • Chip preparation method, DNA or protein immobilization method and chip
  • Chip preparation method, DNA or protein immobilization method and chip
  • Chip preparation method, DNA or protein immobilization method and chip

Examples

Experimental program
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Effect test

Embodiment 1

[0055] In this example, on a glass substrate with epoxy silane on the surface, DNA is immobilized on the surface of the substrate through the amino group of DNA to form the DNA chip of this example. In the preparation process, a weak passivation treatment was added after the fixation treatment, and the surfactant CTAB was used as the catalyst in this example for the weak passivation treatment. This example compares the effect of adding or not adding CTAB to DNA fixation during weak passivation.

[0056] In this example, the “in-channel” fixation method is used to fix DNA on the chip substrate, that is, the chip substrate is packaged first, and then various reaction reagents and washing reagents are respectively introduced into the packaged chip channel by a fluid device to achieve fixation. Chemical reactions such as treatment and passivation treatment. Such as figure 1 As shown, after the chip substrate is packaged, each chip channel is formed independent of each other, and eac...

Embodiment 2

[0068] The DNA chip preparation materials and procedures of this example are the same as those in Example 1, except that during the fixation process, the DNA content in the fixation reaction solution is 0.01nM, 0.04nM, 0.07nM, and 0.1nM, respectively. The remaining conditions are the same as those in the example. One is the same to verify the effect of different concentrations of DNA on the chip.

[0069] The specific preparation method of the DNA chip in this example is as follows:

[0070] (1) Fixation treatment: Pass the fixation reaction solution into the channel of the chip substrate to perform the fixation reaction. The fixation reaction solution in this example is the fixation solution AT-01 containing DNA and 50 thymine deoxynucleotides; DNA containing 5'end with amino modified NH 2 And Cy3 fluorophore modification; AT-01 is composed of Na 2 CO 3 / NaHCO 3 , 0.6mM CTAB, pH 9.78, the volume of the circulating solution is 1mL, the fluid velocity is 1mL / min, the reaction time i...

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Abstract

The invention discloses a chip preparation method, a DNA or protein immobilization method, and a chip. The chip preparation method of the invention comprises performing weak passivation treatment after immobilization treatment, wherein the weak passivation treatment comprises contacting a weak passivation reaction solution containing a catalyst with a chip subjected to immobilization treatment, soas to promote binding of DNA or protein to the surface of a substrate, and to enable the DNA or protein to be sufficiently immobilized on the surface of the substrate. The preparation method of the invention increases the weak passivation treatment step after the immobilization treatment, so that the DNA or protein can be more fully immobilized on the surface of the substrate, the quality and efficiency of DNA or protein immobilization are improved, also DNA or protein is more fully immobilized, the relationship between the amount of the DNA or protein immobilized on the chip and the concentration of the DNA or protein initially added is more closely, and thus the immobilization amount of the DNA or protein is highly controllable, the repeatability is good, and a foundation is laid for preparing high-quality DNA or protein chips.

Description

Technical field [0001] This application relates to the field of chip preparation, in particular to a method for preparing a chip, a method for fixing DNA or protein, and a chip. Background technique [0002] DNA is an important genetic material for many organisms, including humans. It contains a lot of important information and can provide key information for physiological or pathological testing. Efficient and accurate detection of DNA, especially the ability to quantitatively identify changes in base information, not only has great scientific research significance, but also has great clinical application value. DNA chip-based identification and detection methods, compared with other detection methods such as electrophoresis detection, PCR detection or test strip detection, DNA chip detection has the advantages of high detection throughput, good accuracy, convenient detection, and no need for special test personnel. DNA chip is an important platform to realize DNA chip detectio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C40B40/06C40B40/10C12Q1/6837G01N33/68
CPCC40B40/10C40B50/06C40B40/08
Inventor 高锦鸿赵智赵陆洋王琦
Owner GENEMIND BIOSCIENCES CO LTD
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