DNA immobilization method based on nucleic acid immobilization by gel and use thereof

An immobilization method and nucleic acid technology, applied in the field of target nucleic acid detection, can solve problems such as the reduction of immobilization efficiency, and achieve the effects of eliminating possibility, high immobilization efficiency and high loading capacity.
CN101481733AInactive Publication Date: 2009-07-15HANGZHOU ENSHI GENE TECH DEV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
HANGZHOU ENSHI GENE TECH DEV
Publication Date
2009-07-15
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention relates to a method of immobilizing DNA on gel, in particular to a DNA immobilization method based on immobilization of nucleic acid on the gel and the application thereof. The immobilization method of a DNA sample comprises three steps of raw material selection, mixing and polyreaction. In the step of raw material selection, the selected raw materials and dosage are as follows: 10 percent of 10*TBE(pH13), 10-70 percent of 40 percent methacrylamide storage liquid, 1muL-5ml diluted degeneration amino terminal modified nucleic acid, 1-10 percent of N, N, N, N-tetramethyl ethylenediamine, 1-10 percent of 10 percent ammonium persulfate, and water. In the step of mixing, configured gel raw material is added with amino terminal modified nucleic acid to be mixed. The nucleic acid amount in the mixture is 10-1,000ng / 3muL. In the step of polyreaction, 5muL mixture is put in a PCR reaction tube at 37 DEG C for 30min. After the polyreaction is finished, the mixture is molded into gel shape, and the amino terminal modified nucleic acid is immobilized by the polyreaction in the gel. The method has the advantages of high bearing capacity, immobilization of the terminal modified nucleic acid molecule by stable covalent bond formed between the molecule and a solid phase carrier and capacity of enduring dramatic conditions of the PCR reaction, etc.
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Description

technical field

[0001] The invention relates to a method for immobilizing DNA on a gel, which can be used for preparing DNA microarrays, repeated utilization of immobilized DNA, and the like. The invention also relates to a material on which DNA is immobilized according to said method and a method for detecting a target nucleic acid using said material. Background technique

[0002] The integrity of our genetic material is constantly challenged due to constant chemical exposure (environmental substances, food, etc.). Genomic DNA is not static in the process of disease and throughout a person's life, and it is increasingly necessary to study the dynamic changes at the genome level.

[0003] Due to technical bottlenecks, systematic comparative studies on the dynamic changes at the genome level during disease processes are still not possible. However, the need to preserve individual genome DNA for current or future research obviously has extremely important practical signific...

Claims

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