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A kind of dna tetrahedral nanostructure signal probe and its application

A technology of DNA probe and signal probe, applied in the field of DNA tetrahedron nanostructure signal probe, can solve the problem of low efficiency of enzyme reaction

Inactive Publication Date: 2017-02-01
SHANGHAI ADVANCED RES INST CHINESE ACADEMY OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since a single-stranded DNA signaling probe usually only binds to one HRP enzyme, the efficiency of the enzymatic reaction is not high

Method used

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  • A kind of dna tetrahedral nanostructure signal probe and its application
  • A kind of dna tetrahedral nanostructure signal probe and its application
  • A kind of dna tetrahedral nanostructure signal probe and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] Four single-stranded DNAs used to assemble and form tetrahedral DNA nanostructure signal probes: tetra-a-reporter (77bp, molecular weight 23718.43, ssDNA); tetra-b-biotin (55bp, molecular weight 17032.04, 5'-end modified biotin ssDNA ); tetra-c-biotin (55bp, molecular weight 16909.915' end modified biotin ssDNA); tetra-d-biotin (55bp, molecular weight 16888.84, 5'end modified biotin); all synthesized by Shanghai Shenggong Company.

[0082] The four DNAs constituting the tetrahedral nanostructure signal probe contain three domains, each of which is complementary to the corresponding domains of the other three single-stranded DNAs (17 pairs of base complements), and each single-stranded DNA surrounds four sides. One face of the body structure has a circle, and each vertex contains two bases (non-complementary, flexible) to bend. The 3'and 5'ends of the single-stranded DNA converge on the four vertices of the tetrahedron. Tetra-a-reporter extends a DNA sequence at the 5'end a...

Embodiment 2

[0096] The electrochemical detection method is as follows:

[0097] 1) Self-assembled tetrahedral DNA capture probe and tetrahedral DNA nanostructure signal probe

[0098] Capture probe synthesis: Take equal amounts of tetra-a, b, c, d four single-stranded DNA, and use TM buffer (20mMtris, 50mM MgCl 2 , pH8.0) dilute to make the concentration 1uM and the volume 100uL. After 3 minutes at 95 degrees Celsius, the temperature was immediately lowered to 4 degrees Celsius.

[0099] Signal probe synthesis: Take equal amounts of tetra-a-reporter, b-biotin, c-biotin, d-biotin four single-stranded DNA, and use TM buffer (20mM tris, 50mM MgCl 2 , pH8.0) dilute to make the concentration 1uM and the volume 100uL. After 3 minutes at 95 degrees Celsius, the temperature was immediately lowered to 4 degrees Celsius.

[0100] 2) Clean and polish the electrode and assemble

[0101] Take a gold electrode with a diameter of 2mm, first polish it with 0.3um and 0.5um alumina powder in turn, then sonicate it...

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Abstract

The invention provides a DNA tetrahedral nano-structure signal probe. The signal probe is a DNA signal probe having a tetrahedral base, wherein the signal probe is formed by self-assembling single-stranded DNA of a DNA recognition sequence extending from 5' end and single-stranded DNA of three chains with 5' ends modified with labeled molecule; the DNA recognition sequence extends from a vertex of the tetrahedral structure, the DNA recognition sequence is complementarily combined with to-be-detected target DNA or miRNA, and other three vertexes, due to the labeled molecules, are specifically combined with modified signaling modules, so as to achieve multi-signal amplification. The signal probe disclosed by the invention greatly improves the sensitivity of the target DNA or miRNA subjected to electrochemical detection, and a limit of detection can be as low as 1fM.

Description

Technical field [0001] The invention relates to the field of biotechnology, in particular to a DNA tetrahedral nanostructure signal probe and its application. Background technique [0002] DNA electrochemical biosensors have huge potential applications in various fields such as clinical medicine, food inspection, environmental testing, and anti-terrorism due to their advantages of speed, sensitivity, low cost, and easy miniaturization. An important aspect that determines the performance of DNA electrochemical biosensors is the signal amplification system. [0003] As a biomarker, DNA has a very important position in disease diagnosis. Therefore, ultra-sensitive DNA sensors for detecting biomarkers of low concentrations of DNA in clinical samples are essential. Among them, the electrochemical DNA sensor is a promising one because of its fast response speed, convenient use, low cost, and direct DNA detection. The electrochemical DNA sensor was invented for the first time in 2003, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11G01N27/327G01N27/48
Inventor 宓现强曾冬冬
Owner SHANGHAI ADVANCED RES INST CHINESE ACADEMY OF SCI
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