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Three-function magnetic aptamer three-nucleic-acid nano probe

An aptamer and three-function technology, applied in nanotechnology, nanotechnology, nano-optics, etc., achieves the effect of simple construction and simultaneous qualitative and quantitative determination

Active Publication Date: 2021-03-16
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The currently widely used detection methods for exosomes or other ligands are difficult to achieve simultaneous qualitative and quantitative detection of ligand substances, while nucleic acid detection technology has been relatively mature, so the difficult-to-determined ligands are converted into easier-to-detect Single-stranded nucleic acid targets are of great interest

Method used

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  • Three-function magnetic aptamer three-nucleic-acid nano probe

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1 Construction of a three-functional magnetic aptamer triple nucleic acid nanoprobe

[0049] 1. Experimental materials

[0050] HEPES and sodium chloride were purchased from Aladdin, and magnetic beads were purchased from Zhongke Leiming (Beijing) Technology Co., Ltd. All experimental water was from Milli-Q pure water system.

[0051] The sequence design is as follows (SEQ ID NO:1-2):

[0052]

[0053] Note: The underlined part indicates the aptamer sequence of CD63 protein on the surface of exosomes;

[0054] Bold parts indicate the sequences that form the triple nucleic acid.

[0055] 2. Construction and characterization of aptamer triple nucleic acid nanoprobes

[0056] Mix the aptamer three-nucleic acid backbone chain labeled with a fluorescent group and a quencher group at the 5' end and 3' end respectively with the single-stranded DNA target at a ratio of 1:1.1, and incubate in the dark at 37°C for 1 h. Obtain aptamer three nucleic acid nanoprobe s...

Embodiment 2

[0061] Example 2 Verification of Effective Target Conversion Efficiency

[0062] Mix the aptamer three-nucleic acid backbone chain labeled with a fluorescent group and a quencher group at the 5' end and 3' end respectively with the single-stranded DNA target at a ratio of 1:1.1, and incubate in the dark at 37°C for 1 h. Obtain the aptamer three nucleic acid nanoprobe stock solution with a concentration of 200nM; use a microplate reader to detect the fluorescence intensity under the conditions of excitation light 488nm and emission light 500-600nm, after saving the record, record the fluorescence intensity at the time of the maximum emission light 532nm I1, the fluorescence intensity of the control group was recorded as I2, and the fluorescence quenching rate Q was calculated, and the calculation formula was: Q=(I2-I1) / I2×100%.

[0063] Take 50 µL of the obtained stock solution, add 15 µL of exosome solution as the experimental group; take 50 µL of the obtained stock solution, ...

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Abstract

The invention provides a three-functional magnetic aptamer three-nucleic-acid nanoprobe. The three-functional magnetic aptamer three-nucleic-acid nanoprobe comprises magnetic beads (1), an aptamer three-nucleic-acid skeleton chain (2) and a single-stranded DNA target (3). The magnetic aptamer three-nucleic-acid DNA nanoprobe with triple functions is constructed by taking an aptamer sequence as a recognition element, three nucleic acids as a molecular conversion element and magnetic beads as a separation and purification element, and equivalent conversion of ligands and single-stranded DNA is realized. Ligands which are not easy to detect are converted into single-stranded DNA targets which are easy to determine, so that the problem that the ligands are not easy to directly determine qualitatively and quantitatively at the same time at present is solved.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a three-functional magnetic aptamer triple nucleic acid nanoprobe. Background technique [0002] Trinucleic acid belongs to a class of functional nucleic acids, and is one of the tools for constructing new biosensors. Trinucleic acid is a triple-stranded nucleotide combined with a third strand in the form of Hooster hydrogen bonds or anti-Houster hydrogen bonds in the major groove of the double-stranded nucleotide double helix structure. The formation of trinucleic acid is closely related to conditions such as nucleic acid sequence, base composition, ion environment, and pH value. Due to the reversibility of its structure, trinucleic acid can be used as a "switch" in biosensors, and is often used for target substance conversion. [0003] The currently widely used detection methods for exosomes or other ligands are difficult to achieve simultaneous quali...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C09K11/06C09K11/02G01N21/64B82Y20/00G01N33/569G01N33/58
CPCB82Y20/00C09K11/025C09K11/06G01N21/6428G01N33/56966G01N33/582G01N2021/6432
Inventor 许文涛黄昆仑王馨娴李舒婷程楠朱龙佼
Owner CHINA AGRI UNIV
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