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A single quantum dot fluorescent nanosensor based on enzyme-free catalyzed self-assembly and its preparation method and application

A fluorescent nanometer, non-enzyme-catalyzed technology, applied in the field of biological analysis, can solve the problems of reducing detection accuracy, complicating the design of the scheme, increasing the detection cost, etc., and achieves the advantages of less sample consumption, improved sensitivity, and reduced false positive signals Effect

Active Publication Date: 2022-05-27
SHANDONG NORMAL UNIV
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Problems solved by technology

However, the use of enzyme-catalyzed signal amplification by these methods will inevitably increase assay cost, complicate protocol design, and reduce assay accuracy

Method used

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  • A single quantum dot fluorescent nanosensor based on enzyme-free catalyzed self-assembly and its preparation method and application
  • A single quantum dot fluorescent nanosensor based on enzyme-free catalyzed self-assembly and its preparation method and application
  • A single quantum dot fluorescent nanosensor based on enzyme-free catalyzed self-assembly and its preparation method and application

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Embodiment 1

[0045] Detection of circulating microRNAs and formation of QD / DNA / Cy5 nanocomplexes: The detection of circulating microRNAs consists of two sequential steps: first, signal probes and capture probes were mixed in 1× reaction buffer (20 mM per liter of triplicates). hydroxymethylaminomethane-hydrochloric acid, 140 mM NaCl, 5 mM KCl, pH 7.5) diluted to 2 μM per liter, then incubated at 95°C for 5 min, and Slowly cool to room temperature to form hairpin structure. The prepared hairpin probes were used immediately or kept at -20°C for later use. Second, the 60 μl reaction system includes the indicated concentration of miR-21, 5.4 μl capture probe (2 μmol per liter), 5.4 μl signal probe (2 μmol per liter), 6 μl 10× reaction Buffer (200 mmol / L Tris-HCl, 1.4 mmol / L NaCl, 50 mmol / L KCl, pH 7.5, react at 37°C for 3 hours, then add 6 Microliters of 605QDs (50 nanomoles per liter) were incubated at 37°C for 10 minutes to form 605QD / DNA / Cy5 nanostructures.

[0046] Polyacrylamide gel el...

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Abstract

The invention relates to a single quantum dot fluorescent nanosensor based on non-enzyme catalyzed self-assembly and its preparation method and application. It consists of two hairpin DNA probes and quantum dots modified by streptavidin protein, one of which is The DNA probe is a biotin-labeled capture probe, the other hairpin DNA probe is a signal probe labeled with fluorophore cyanine 5, and the two hairpin DNA probes are assembled on streptavidin-modified quantum dots (SA‑QDs) surface to form single quantum dot fluorescent nanosensors. It has applications in the detection of endogenous microRNA in cancer cells and circulating microRNA in patient blood samples with high selectivity.

Description

technical field [0001] The invention belongs to the technical field of biological analysis, and in particular relates to a single quantum dot fluorescent nano-sensor based on enzyme-free catalytic self-assembly and a preparation method and application thereof. Background technique [0002] The information disclosed in this Background section is only for enhancement of understanding of the general background of the invention and should not necessarily be taken as an acknowledgement or any form of suggestion that this information forms the prior art already known to a person of ordinary skill in the art. [0003] MicroRNAs (microRNAs) are small, single-stranded, non-protein-coding RNAs that are widely found in plants and animals. As post-transcriptional regulators of gene expression, microRNAs are involved in a variety of normal physiological processes such as cell proliferation, differentiation and death. Meanwhile, abnormal expression levels of microRNAs are closely related...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
CPCG01N21/6402G01N21/6486
Inventor 张春阳马飞张倩
Owner SHANDONG NORMAL UNIV
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