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Support for fixing nucleotide and process for producing the same

A technology for nucleotides and oligonucleotides, which is applied to the support phase for immobilizing nucleotides and its preparation, and can solve problems such as dealing with damage

Inactive Publication Date: 2003-02-12
TOYO KOHAN CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

However, when the target DNA is directly bound to these chemically modified sites by an amide bond, the end of the target DNA is in danger of being damaged by the treatment during, for example, the chemical bonding process.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Preparation of support phase for immobilizing nucleotides with EcoRI restriction site

[0048] (1) Chemical modification of substrate

[0049] The substrate containing diamond is placed in chlorine gas and irradiated with ultraviolet light to chlorinate the surface of the substrate. Afterwards, it is placed in ammonia gas and irradiated with ultraviolet rays to make it ammoniated, and then the substrate is carboxylated by refluxing acid chloride in chloroform, so that the surface of the substrate is chemically modified.

[0050] (2) Immobilization of single-stranded oligonucleotides

[0051] The chemically modified matrix prepared in (1) above was immersed in 1,4-dioxane solution for 15 minutes (the ratio of diamond matrix and dioxane was 1:100 μl), and 2.5 mg / ml carbodiimide and 1.5mg / ml N-hydroxysuccinimide to dehydrate and condense the terminal carboxyl group. After the reaction is complete, the product is washed with water, then with 1,4-dioxane solution, and th...

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Abstract

A support for fixing a nucleotide which contributes to the efficient clarification of DNA without damaging the terminal parts of the DNA and, therefore, is useful in the fields of molecular biology, biochemistry and the like. This support for fixing a nucleotide is a chemically modified substrate on which oligonucleotides are immobilized characterized in that the oligonucleotides have a restriction enzyme cleavage site. This support is produced by chemically modifying the substrate, immobilizing a single-stranded oligonucleotide thereon, then hybridizing the single-stranded oligonucleotide with another single-stranded oligonucleotide having a base sequence complementary thereto, and ligating an oligonucleotide having a restriction enzyme site at the end thereof.

Description

technical field [0001] The invention relates to a support phase for immobilizing nucleotides and a preparation method thereof. The support phase can be used for immobilizing nucleic acids and the like, and can be applied to the related fields of molecular biology and genetic engineering. Background of the invention [0002] Gene analysis is widely used in the fields of molecular biology and genetic engineering, and has also been used in medical fields such as disease diagnosis in recent years. [0003] In gene analysis, DNA chips have been developed in recent years, and the speed of gene analysis is also getting faster and faster. The traditional DNA chip technology is to apply macromolecular substances such as polylysine to the surface of glass slides or silicone substrates, and then fix DNA. Another method is to synthesize oligonucleotides on the surface of glass substrates using semiconductor techniques such as photolithography. [0004] However, i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/00C07H21/00C12N15/10C12Q1/68G01N33/53G01N37/00
CPCC07B2200/11C07H21/00C12N15/1006C12Q1/6837C12Q2521/501C12Q2521/301C12Q1/68
Inventor 丹花通文冈村浩高木研一高桥浩二郎
Owner TOYO KOHAN CO LTD
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