Multi-copy monomolecular nucleic acid array chip

A molecular nucleic acid and microarray chip technology, which is applied in the field of biomedical single-molecule nucleic acid array chip manufacturing, can solve the problem of increasing the cost of the chip and the complexity of the preparation process, hindering the use of microarray chip technology, and affecting the accuracy and reliability of the chip and other problems, to achieve the effect of short preparation time, low cost and high preparation cost

Inactive Publication Date: 2006-12-13
SOUTHEAST UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

On the one hand, this limits the further improvement of chip probe density; on the other hand, it greatly increases the cost of the chip and the complexity of the preparation process; at the same time, the purity and immobilization efficiency of sample probes will also affect the accuracy and reliability of chip detection. sex
These factors have greatly hindered the use of microarray chip technology, especially for the use of high-throughput DNA chips

Method used

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  • Multi-copy monomolecular nucleic acid array chip
  • Multi-copy monomolecular nucleic acid array chip
  • Multi-copy monomolecular nucleic acid array chip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1. Application of rolling circle amplification technique to prepare single-molecule multi-copy nucleic acid microarray chip.

[0034] ●Selection of enzyme cutting sites and preparation of circular single-stranded genomic DNA fragments

[0035] Restriction enzyme digestion of genomic DNA: firstly, select an appropriate endonuclease (as shown in Table 1) to digest genomic DNA by bioinformatics method, and cut the genome into cohesive end fragments with a size of 50-200 bp. At the same time, according to the characteristics of the end sequence after each endonuclease digestion, design 2 universal linkers. image 3 Taking the combination of Nla III endonuclease and Dpn II endonuclease as an example, the design diagram of the two universal linker sequences. After the human genome DNA extracted from the blood was digested under different restriction endonucleases, 1 μl of the product was electrophoresed on 1.5% agarose gel for 30 minutes to detect the digestion effec...

Embodiment 2

[0043] Example 2, Application of microemulsion technology to prepare single-molecule multi-copy nucleic acid microarray chip

[0044] Microemulsion means that one or several liquids are dispersed in another immiscible liquid in the form of particles (droplets or liquid crystals) to form a multiphase dispersion system with considerable stability. Because their appearance is often milky, so called an emulsion. Its main features are thermodynamic stability, isotropy, transparent or translucent appearance, uniform particle size from nanometer to micron level. Microemulsion related technology is very common in chemistry. We use its thermodynamic stability and other characteristics to place biochemical reactions such as PCR amplification in microemulsion droplets, so that the reaction can be carried out efficiently and quickly.

Embodiment approach

[0045] Concrete implementation scheme is as follows:

[0046] ●Single-molecule parallel PCR in microemulsions

[0047] The preparation of water-in-oil microemulsion at first selects suitable oil phase (mineral oil etc.) and surfactant (Tween, SDS, Triton etc.), is water phase with parallel PCR amplification system, oil phase and water phase by 20: Mix at a ratio of 1 to 1:1, and prepare microemulsions by shaking or ultrasonication. The particle size and uniformity of the prepared microemulsion were observed under a microscope. By controlling the ratio of the water phase and the oil phase, the preparation method, the preparation temperature, the concentration of the surfactant and other conditions, a microemulsion system with uniform particle size can be obtained, and the particle size of the prepared microemulsion can be further optimized to be suitable for polymer molecular microemulsion. Basic single-molecule multi-copy amplification system.

[0048] Parallel PCR amplific...

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Abstract

The invention relates to a new monomolecular nucleotide microarray chip with high density and distinguishability. A plurality of polymer high molecule containing nucleotide fragment of this chip can randomly fix on a solid base-plate and can form a monomolecular nucleotide microarray with high density, and can make every array point contain one polymer high molecule, in which the said polymer high molecule contains some similar copies of nucleotide fragment. This chip can be assembled to many kinds of chips used for functional genom study, testing order of complete gene of special types, gene transferring and expression efficiency testing, and testing of kinds of genom DNA modification chart(such as DNA methylation chart). This chip will have great value in the fields of molecular mechanism study in human being life process, genom DNA information decoding, individualized medicine, disease predicting and precaution, medicine developing and individuation.

Description

technical field [0001] The invention relates to a high-density nucleic acid microarray chip, which can be used for functional genome research, such as genome sequencing of specific species, detection of gene transcription and expression profiles, and various genome DNA modification profiles (such as DNA methylation profile ) detection, belonging to the technical field of biomedical single-molecule nucleic acid array chip manufacturing. Background technique [0002] The human genome (sequencing) project has been completed, and the post-genome project has entered into implementation. Especially after the completion of the Human Genome Project, relevant nucleic acid sequence data has grown exponentially, and more and more genome sequences of animals, plants, and microorganisms have been determined, providing a wealth of information resources for comparative genomics research. How to analyze and compare a large amount of biomolecular information, find common molecular mechanism...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6837
Inventor 陆祖宏白云飞葛芹玉
Owner SOUTHEAST UNIV
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