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Solid phase DNA chip hybridization solution and hybridization method

A DNA chip and hybridization method technology, which is applied in the field of solid-phase DNA chip hybridization solution and hybridization, can solve the problems of spots in hybridization results, influence chip quality, poor hybridization signal, etc., and achieve shortened hybridization time, easy storage, and improved hybridization. effect of the signal

Inactive Publication Date: 2011-01-19
TIANJIN BIOCHIP TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Low hybridization temperature often leads to poor hybridization signal, high background signal, spots in hybridization results, etc., which affects the quality of the chip

Method used

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  • Solid phase DNA chip hybridization solution and hybridization method
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  • Solid phase DNA chip hybridization solution and hybridization method

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Embodiment 1

[0045] The composition of the hybridization solution: NaCl 717mM, Na-MES 400mM, EDTA 100μM, SDS 0.2% (w / v). Hybridization method of hybridization solution:

[0046] 1) When the PCR amplification product is single-stranded labeled with fluorescein, the PCR labeled product is dried at 65°C;

[0047] 2) Redissolve the labeled product with 10 μl preheated deionized water, then add 10 μl preheated hybridization solution, and mix well;

[0048] 3) The mixed solution was added to the incubation chamber, and the hybridization temperature was probe Tm-10°C, and the hybridization chamber was closed and placed in a water bath; the temperature was controlled at 50°C, and the hybridization time was 6 hours.

[0049] The hybridization liquid described in the present invention is a 2× hybridization liquid, which should be diluted 1:1 with deionized water when used. The hybridization solution and deionized water should be fully preheated at 65°C before dissolving the PCR product. The stora...

Embodiment 2

[0051] The composition of the hybridization solution: NaCl 717mM, Na-MES 400mM, EDTA 100μM, SDS 0.2% (w / v).

[0052] Hybridization method of hybridization solution:

[0053] 1) When the PCR amplification product is single-stranded labeled with fluorescein, the PCR labeled product is dried at 65°C;

[0054] 2) Redissolve the labeled product with 10 μl preheated deionized water, then add 10 μl preheated hybridization solution, and mix well;

[0055] 3) The mixed solution is added to the incubation chamber, the hybridization temperature is probe Tm-10°C, and the hybridization chamber is closed and placed in a water bath; the temperature is controlled at 60°C, and the hybridization time is 4 hours.

[0056] The hybridization liquid described in the present invention is a 2× hybridization liquid, which should be diluted 1:1 with deionized water when used. The hybridization solution and deionized water should be fully preheated at 65°C before dissolving the PCR product.

Embodiment 3

[0058] The composition of the hybridization solution: NaCl 717mM, Na-MES 400mM, EDTA 100μM, SDS 0.2% (w / v).

[0059] Hybridization method of hybridization solution:

[0060] 1) When PCR amplification is performed while labeling, the PCR product is fully annealed at 95°C;

[0061] 2) Put the annealed PCR product on ice quickly, and place it for 10 minutes to quench it;

[0062] 3) Mix the quenched PCR product with 10 μl deionized water and 10 μl fully preheated hybridization solution;

[0063] 4) Add the mixed solution into the incubation chamber, close the hybridization chamber, and perform hybridization in a water bath.

[0064] The hybridization liquid described in the present invention is a 2× hybridization liquid, which should be diluted 1:1 with deionized water when used, and the storage temperature of the hybridization liquid is 4°C or -20°C.

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Abstract

The invention relates to a solid phase DNA chip hybridization solution and a hybridization method. The solid phase DNA chip hybridization solution is composed of raw materials of NaCl, Na-MES, EDTA and SDS, wherein NaCl is 717mm, Na-MES is 400mm, EDTA is 100 mu m, and SDS accounts for 0.2%(w / t). The main ingredients of the hybridization solution are as follows: NaCl, Na-MES, EDTA and SDS. During hybridization, a marked PCR product is only dried and is mixed and dissolved by preheated hybridization solution and deionized water at the ratio of 1:1, and the obtained mixture is added into an incubation chamber to be incubated. The method of the invention is simple and easy to realize, can improve hybridization temperature and can effectively shorten hybridization time. The hybridization solution can preferably control a background value generated by hybridization, improves hybridization signal, reduces interference of the background value to the signal value and optimizes the hybridization effect.

Description

technical field [0001] The invention relates to a solid-phase DNA chip hybridization solution and a simple hybridization method. Background technique [0002] With the gradual implementation of the Human Genome Project and the rapid development of molecular biology-related disciplines, more and more genome sequences of animals, plants, and microorganisms have been determined, and gene sequence data is growing rapidly at an unprecedented rate. . Gene chip (also known as DNA chip, biochip) technology is the product of this scientific development requirement, and its appearance provides a bright prospect for solving such problems. Gene chip technology has been increasingly used in gene diagnosis, analysis, and gene therapy. This technique refers to immobilizing a large number of probe molecules (usually higher than 400 per square centimeter lattice density) on the support and hybridizing with the labeled sample molecules, and then obtaining the sample molecules by detecting t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 王磊张恋心杨磊李荣荣冯露曹勃阳
Owner TIANJIN BIOCHIP TECH CO LTD
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