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Method for identifying specific DNA sequences in germplasm resources based on surface plasma resonance technology

A DNA sequence and surface plasma technology, applied in the field of specific DNA sequence identification in germplasm resources, can solve the problems of high cost and large amount of consumables, and achieve the effects of prolonging service life, reducing detection cost and easy regeneration

Inactive Publication Date: 2012-09-12
王利兵
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AI Technical Summary

Problems solved by technology

[0004] In order to solve the problems of large amount of consumables and high cost in the existing specific DNA sequence analysis technology, and at the same time meet the identification requirements of various DNA sequences in germplasm resources, the present invention provides a surface plasmon of specific DNA in germplasm resources Resonance identification method

Method used

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  • Method for identifying specific DNA sequences in germplasm resources based on surface plasma resonance technology
  • Method for identifying specific DNA sequences in germplasm resources based on surface plasma resonance technology

Examples

Experimental program
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Effect test

Embodiment

[0028] Embodiment: select a section of SSR sequence of the rice dominant early-maturing gene Ehd as the identification object, and the specific operation steps are as follows:

[0029] (1) Aptamer immobilization on the surface of the sensor: select the sensor chip modified by streptavidin SA, pass 20 μL of biotin-labeled immunoglobulin E aptamer with a concentration of 30 μg / L, on the surface plasmon The immunoglobulin E aptamer was immobilized on the sensor chip;

[0030] (2) Coupling of the specific SSR sequence of rice dominant early-maturing gene Ehd and immunoglobulin E: react the specific SSR single chain with p-phenylene diisothiocyanate, and then couple with the amino group of immunoglobulin E to form a p-phenylene diisothiocyanate-linked immunoglobulin E-specific SSR sequence conjugate, ready for use;

[0031] (3) Preparation of standard solution: Use 0.02mol / L (pH=7.4) HEPBS buffer to prepare a specific single-stranded DNA solution labeled with immunoglobulin E, wit...

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Abstract

The invention discloses a method for identifying specific DNA sequences in germplasm resources based on surface plasma resonance technology, belonging to the technical field of germplasm resource identification. The method comprises four steps as the followings: fixing an aptamer on the surface of a sensing surface, marking a sample, identifying the sample, and regenerating a sensing element. The method utilizes the same sensing chip to identify different DNA sequences. The method is characterized by using immunoglobulin E as a marked molecular probe, and connecting the sensing chip surface with the immunoglobulin E aptamer; carrying out a series of pretreatment on a food sample, adding a single-stranded DNA immunoglobulin E marker, after mixing, competitively binding a corresponding aptamer of the single-stranded DNA modified by metal nano particles; detecting the content of immunoglobulin E by using a surface plasma resonance detector, indirectly identifying the specific single-stranded DNA sequences; and regenerating the sensing chip for detecting the other specific DNA sequences. The invention provides an effective identification method of specific DNA sequences in germplasm resources, thereby realizing the identification of different DNA sequences by the same surface plasma resonance sensing chip, and the method has extremely high versatility.

Description

technical field [0001] The invention relates to a specific DNA sequence identification technology in germplasm resources, especially based on surface plasmon resonance technology, nucleic acid aptamers specifically compete with unlabeled and labeled samples for identification, and indirect identification using immunoglobulin E-modified sensing elements Methods for specifying DNA sequences in germplasm. Background technique [0002] Germplasm resources are the basic strategic resources for the sustainable development of the national economy. However, in recent years, my country's germplasm resources have been seriously lost. On the one hand, my country's human genetic resources, important economic crop resources, medicinal resources, and industrial microbial resources have been lost to foreign countries through illegal means; my country has established a base for rough processing of plant extracts, and predatoryly imported high-quality medicinal materials and resource extract...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/55G01N21/552
Inventor 王利兵于艳军苏荣欣
Owner 王利兵
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