High efficiency full genome fixing method

A whole genome, fixed method technology, applied in the field of genome detection in biomedicine, can solve the problems of limited detection sites and incomplete information, and achieve the effects of saving detection costs, accurate information, and saving preparation time

Inactive Publication Date: 2007-04-11
SOUTHEAST UNIV
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  • Claims
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AI Technical Summary

Problems solved by technology

Although these technologies can complete the detection of mutations or nucleotide polymorphisms to a certain extent, they first need to amplify the target fragments to improve the sensitivity of detection, so the sites they detect are quite limited, so the information obtained is incomplete

Method used

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  • High efficiency full genome fixing method

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example 1

[0033] Example 1 Yeast whole genome and preparation method of whole genome DNA chip

[0034] Referring to accompanying drawing 1, on a flat glass slide, process by photolithography into the microporous plate of diameter 50 microns, depth 50 microns, then use dichlorodimethylsilane to carry out hydrophobic treatment on the surface of microporous plate. After the micropores were washed with water and dried, they were modified with 5% 3-aminopropyltrimethoxysilaneacetone for 1 hour, and treated with 5% glutaraldehyde for 2 hours, and then reacted with the oligonucleotide solution modified with the amino group at the 5' end. The oligonucleotides are immobilized in the microwells. On the other hand, the yeast genome is cut into fragments with a size of about 500 bases by enzymes, and these fragmented nucleic acid sequences are connected with a pair of universal linkers under the action of ligase, one of which is connected to the microwell The oligonucleotide molecular sequence imm...

example 2

[0037] Example 2 The preparation method of human whole genome and its whole genome DNA chip

[0038] Referring to accompanying drawing 1, a microporous plate with a diameter of 50 microns and a depth of 50 microns is processed by photolithography on a flat glass sheet, and then the surface of the microporous plate is subjected to hydrophobic treatment with dichlorodimethylsilane. Mix oligonucleotides modified with acrylamide groups at the 5' end with acrylamide monomers, ammonium persulfate, etc., and fill them in micropores, and let the water in the prepolymer volatilize naturally. When the water volatilizes to the prepolymer When only two-thirds to one-third of the micropores are occupied, the glass sheet is placed under vacuum, and the polymerization reaction is promoted to form a gel under the vaporized atmosphere of the initiator and fixed in the micropores.

[0039] On the one hand, the human genome is cut into fragments with a size of 50-1000 bases with enzymes, and the...

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Abstract

The high efficiency full genome fixing method relates to method of amplifying full genome in high efficiency and preparing full genome DNA chip. The method includes: 1. machining pores in one flat hard material and fixing same oligonucleotide molecules; 2. cutting genome nucleic acid with enzyme into 50-1000 base size fragments and connecting the genome fragment nucleotide sequences with one pair of common connexons under the action of enzyme to form genome fragment nucleotide sequences containing connexons; 3. heat denaturing and diluting the genome fragment nucleotide sequences containing connexons and adding to the pores; and 4. adding amplifying reaction liquid into the pores and amplifying the genome fragment nucleotide sequences containing connexons to realize exponential amplification of the full genome sequence in fragment form.

Description

technical field [0001] The invention relates to a high-efficiency whole-genome amplification and a method for preparing a whole-genome DNA chip, belonging to the technical field of genome detection in biomedicine. Background technique [0002] With the deepening of genome research, it will become possible to understand the differences in life, the rules of disease occurrence and development, and the interaction between drugs and living organisms at the genetic level. Although numerous factors contribute to the occurrence of disease, genetic mutations (single nucleotide polymorphisms, methylation, etc.) are widely recognized as an important intrinsic factor. In terms of basic research, gene mutation sites are helpful for the precise positioning of the genome, the study of the genetic law of disease genes, and the cloning of disease-causing genes; Cancer, diabetes, cardiovascular disease, depression, asthma, etc. are affected by many genes and environmental factors. Through l...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12P19/34
Inventor 肖鹏峰
Owner SOUTHEAST UNIV
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