Micro array preparing DNA assembling process

A DNA sequence and microarray technology, applied in the field of DNA assembly and preparation of microarrays, can solve problems such as difficult large-scale ordered assembly and difficult formation of structurally regular microarrays

Inactive Publication Date: 2007-06-27
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] However, there are still problems about the assembly of guanine-rich DNA: whether it is G-wire or Frayed-wire assembly in the literature, the size is about 10nm-1-1μm, it is difficult to control the formation of large-scale ordered assembly, and it is difficult to Difficult to form regular microarrays

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Embodiment 1

[0023] A method for preparing a microarray by DNA assembly, comprising the following steps:

[0024] (1) DNA sequence G 3 (A 2 G 3 ) 3 Dissolve in pure water so that the concentration of the DNA aqueous solution is 10mM, mix the DNA aqueous solution with methanol at a ratio of 3:97 by volume, add the aqueous solution of lithium chloride, and make Li + The final concentration in the solution is 1000mM;

[0025] (2) Warm up the mixed solution to 95°C, naturally cool to 30°C, and incubate at 25°C for 10 hours;

[0026] (3) Soak the mica substrate in the solution prepared in step (2) for 50 minutes, take it out, and dry it naturally to prepare a DNA assembly preparation microarray.

[0027] The G is guanine deoxynucleotide, and A is adenine deoxynucleotide.

[0028] The preparation method of the DNA sequence is not limited, and it can be synthesized by an automatic synthesizer or a phosphoramidate method.

Embodiment 2

[0030] A method for preparing a microarray by DNA assembly, comprising the following steps:

[0031] (1) DNA sequence G 5 (A 3 G 6 ) 2 Dissolve in pure water so that the concentration of the DNA aqueous solution is 1 mM, mix the DNA aqueous solution with ethanol at a volume ratio of 1:99, add the aqueous ammonium chloride solution, NH 4 + The final concentration in the solution is 500mM;

[0032] (2) Warm up the mixed solution to 80°C, naturally cool to 20°C, and incubate at 2°C for 24 hours;

[0033] (3) Dip the monocrystalline silicon substrate in the solution prepared in step (2) for 10 minutes, take it out, and blow it dry with nitrogen to prepare a DNA assembly preparation microarray.

[0034] The G is guanine deoxynucleotide, and A is adenine deoxynucleotide.

[0035] The preparation method of the DNA sequence is not limited, and it can be synthesized by an automatic synthesizer or a phosphoramidate method.

Embodiment 3

[0037] A method for preparing a microarray by DNA assembly, comprising the following steps:

[0038] (1) DNA sequence G 2 (T 5 G 2 ) 5 Dissolve in pure water so that the concentration of the aqueous solution of DNA is 300mM, mix the aqueous solution of DNA with isopropanol in a ratio of 15:85 by volume, add the aqueous solution of sodium chloride, Na + The final concentration in solution is 100 mM;

[0039] (2) Warm up the mixed solution to 85°C, naturally cool to 25°C, and incubate at 20°C for 15 hours;

[0040] (3) Soak the glass substrate in the solution prepared in step (2) for 30 minutes, take it out, and dry it with air to prepare a DNA assembly preparation microarray.

[0041] The G is guanine deoxynucleotide, and T is thymidine deoxynucleotide.

[0042] The preparation method of the DNA sequence is not limited, and it can be synthesized by an automatic synthesizer or a phosphoramidate method.

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Abstract

The micro array preparing DNA assembling process includes the following steps: 1. dissolving DNA sequence in pure water, mixing with low molecular weight alcohol or ketone, and adding water solution containing Li+, NH4+, Na+ or K+; 2. heating to 80-95 deg.c, cooling to 20-30 deg.c, and incubating at 2-25 deg.c for 10-24 hr; and 3. soaking mica substrate in the solution obtained in the last step for 10-50 min, taking out and drying to obtain DNA assembling micro array. The preparation process is simple, and the prepared DNA assembling micro array has branched structure capable of being regulated through selecting DNA sequence and solution metal particle. The present invention can promote the development of DNA molecular device, DNA circuit, micro channel manufacture, etc. and the DNA assembling micro array may be used as new regular template for guiding the preparation of functional device or material.

Description

technical field [0001] The invention belongs to the technical field of manufacturing functional devices by using a chemical self-assembly method, in particular to a method for preparing a microarray by DNA assembly. Background technique [0002] The unique molecular recognition and assembly properties of deoxynucleic acid (DNA) have attracted extensive attention. The existence form of DNA is not limited to the double helix structure (the DNA double helix structure has a diameter of 2nm and a pitch of 3.4nm), some DNA sequences can also form specific structures such as triplex (Triplex), quadruplex (Quadruplex) or Holliday cross . Through the synergistic interaction between DNA or between DNA and other building units, combined with the characteristics of molecular recognition and self-assembly, the methods and scope of building nano-devices with DNA molecules are enriched. [0003] Under specific ionic strength and pH conditions, guanine (G)-rich DNA forms Hoogsteen pairing...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 李张金利杨薇郑琳
Owner TIANJIN UNIV
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