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Method for detecting microRNA based on triple helix DNA structure

A structure detection and triple helix technology, applied in the field of fluorescent microRNA detection, can solve the problems of time-consuming and large number of samples, and achieve the effect of fast response and simple operation.

Inactive Publication Date: 2017-11-17
UNIV OF JINAN
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Problems solved by technology

[0004] At present, the methods for detecting microRNA mainly include Northern blot analysis, microarray analysis and quantitative real-time PCR, but these methods are not only time-consuming but also require a large number of samples. In order to solve these problems, we have developed a detection microRNA based on triple-helix DNA structure This method can not only quickly detect microRNA, but also is simple to operate, saves time and effort, and can greatly improve the sensitivity of microRNA

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  • Method for detecting microRNA based on triple helix DNA structure
  • Method for detecting microRNA based on triple helix DNA structure

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Embodiment 1

[0028] 1. a method for detecting microRNA based on triple helix DNA structure, is characterized in that comprising the following steps:

[0029] 1.1 Use Adobe illustrator CS4 to design the hydrophobic wax batch printing pattern of the microfluidic paper chip;

[0030] 1.2 Cut the paper into a commonly used A4 size paper, print the hydrophobic wax batch printing pattern designed in step (1.1) onto the A4 paper, and then place the A4 paper with the wax pattern in a flat heater or an oven at 150 ºC heating for 2 min; the wax melts and soaks through the entire thickness of the paper to form a hydrophobic wall;

[0031] 1.3 Probe pre-denaturation

[0032] First, capture probes, auxiliary probes and fluorescent probes were incubated at 95 ºC for 5 min, and the three solutions were naturally cooled at room temperature for 30 min;

[0033] 1.4 Construction of triple helix DNA structure

[0034] Immobilize 80 nM sulfhydryl-labeled capture probes on the surface of the microfluidic pa...

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Abstract

The invention discloses a method for detecting microRNA based on a triple-helix DNA structure, and relates to the technical field of fluorescent microRNA detection. A new triple-helix DNA structure is formed through the hybridization reaction between the three probes on the surface of the microfluidic paper chip, and the triple-helix DNA structure is cut into two parts under the action of endonuclease and exonuclease, One part can circulate and participate in the reaction to achieve signal amplification, and the other part reacts with the target microRNA to generate DNA-RNA double strands, and transfers the formed DNA-RNA double strands to the surface of another paper chip modified with MnO2 nanosheets and adds DSN to detect microRNA. Compared with traditional methods, this method has higher sensitivity, simple operation, and less time-consuming, which greatly promotes the development of a simple and rapid detection system, and helps us to quantitatively analyze microRNA and understand the physiological and pathological mechanisms of organisms. And finally provide new ideas and theoretical basis for the diagnosis and treatment of diseases.

Description

technical field [0001] The invention relates to a method for detecting microRNA based on a triple-helix DNA structure, and belongs to the technical field of fluorescent microRNA detection. Background technique [0002] MicroRNA is currently a very active research field. With the understanding and understanding of microRNA, more and more studies have realized that it is closely related to the formation of tumors. For example, the decreased expression of microRNAs that inhibit oncogenes can lead to the overexpression of oncogenes; while the overexpression of microRNAs that inhibit tumor suppressor genes can lead to decreased expression of tumor suppressor genes. Both overexpression of oncogenes and decreased expression of tumor suppressor genes can lead to tumorigenesis. [0003] MicroRNA is regarded as a novel biomarker and a potential therapeutic target for various diseases. Recent studies have pointed out that microRNA plays an important role in biological processes such ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6837
Inventor 田甜刘海云颜梅于京华葛慎光张彦李丽
Owner UNIV OF JINAN
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