DNA (deoxyribonucleic acid) sequencing method based on micro emulsion isothermal amplification on biochip

A DNA sequencing and biochip technology, applied in the field of biomedicine, can solve problems such as restricting development, taking a long time, and failing to meet the requirements of life science and medical development, and achieve effective, simple and efficient implementation

Inactive Publication Date: 2013-02-13
HANGZHOU DIANZI UNIV
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

Existing DNA detection mainly uses PCR amplification products as templates, which takes a long time and has cumbersome steps, which cannot meet the requirements of life science and medical development, and also restricts the development of the market

Method used

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  • DNA (deoxyribonucleic acid) sequencing method based on micro emulsion isothermal amplification on biochip

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Embodiment Construction

[0017] The present invention will be further described below in conjunction with the accompanying drawings.

[0018] Such as figure 1 As shown, the steps of this embodiment are as follows:

[0019] 1. Modify the surface of the glass slide: modify the surface of the glass slide with aldehyde groups, wash the slide with soapy water, soak in 1:1 methanol and hydrochloric acid for 30 minutes, wash with double distilled water and soak in sulfuric acid for 30 minutes, and then use Wash with double distilled water and 95% ethanol. For the cleaned slides, soak them in 49:1 silane and 95% ethanol for 30 minutes, wash and dry them with ethanol and double distilled water, dry them in an oven at 110 degrees for 30 minutes, wash them with 95% ethanol and double distilled water, and dry them. , soaked in glutaraldehyde to 0.01PB 4:1 ratio for 2 hours, washed with 0.01PB, washed twice with 95% ethanol, and finally washed four times with double distilled water.

[0020] 2. Preparation of a...

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Abstract

The invention discloses a DNA (deoxyribonucleic acid) sequencing method based on micro emulsion isothermal amplification on a biochip. The method comprises the following steps of: firstly, modifying the surface of a slide, performing corresponding aldehyde group, amino group or streptavidin modification on a DNA fragment to be detected; then amplifying the mixture of the modified DNA fragment to be detected and T7gp4 primase in a micro emulsion; then fixing the amplified product to the modified slide for providing a template for sequencing; and finally, performing on-chip extension sequencingon the template fixed to a biochip. In the invention, micro emulsion isothermal amplification and biochip technology are combined, so that large-scale sequencing can be performed on DNA quickly with high flux and high sensitivity.

Description

technical field [0001] The invention is a biochip-based DNA sequencing method, which belongs to the biological detection method in the field of biomedicine. Background technique [0002] The acquisition of all human gene sequences is helpful to understand the pathogenic mechanism of many genetic diseases and cancers, and provides a theoretical basis for new methods such as individual disease diagnosis and gene therapy. And according to the sequence of the genome, we can understand the differences in each person's DNA sequence and the resistance of different individuals to diseases, and carry out corresponding treatments according to each person's gene sequence and characteristics. According to the genome sequence, it can also prevent the possible diseases of the individual, which is of great help to the preventive diagnosis of the disease. The acquisition of the whole human genome sequence depends on the development of DNA sequencing technology. At present, it takes about t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 高力厉力华李宏应南娇王晟黄爱爱徐伟栋刘伟郭晓冬
Owner HANGZHOU DIANZI UNIV
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