Method related to DNA (Deoxyribose Nucleic Acid) folded paper and structure and application thereof

A kind of origami and multi-purpose technology, applied in the field of detection, can solve the problem of low sensitivity

Inactive Publication Date: 2012-07-11
SHANGHAI INST OF APPLIED PHYSICS - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention aims at the deficiency of low sensitivity of the existing methods for detecting target substances, and provides a method involving DNA origami and its structure and application

Method used

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  • Method related to DNA (Deoxyribose Nucleic Acid) folded paper and structure and application thereof
  • Method related to DNA (Deoxyribose Nucleic Acid) folded paper and structure and application thereof
  • Method related to DNA (Deoxyribose Nucleic Acid) folded paper and structure and application thereof

Examples

Experimental program
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preparation example Construction

[0050] In the method for preparing the single-chain staple of the end-modified reactant I described in step 1) of the present invention, the method of connecting the reactant I to the end of the staple chain adopts the prior art. For example, staple strands (unmodified biotin) and biotin-modified single strands of staples are both commercially available. It can also be joined by chemical methods. Other kinds of reactant I also staple the 5' end of the single-stranded nucleotide according to the prior art.

[0051] In step 2) of the present invention, the staple chain and scaffold chain obtained in step 1) are self-assembled, and the scaffold chain and staple single chain (including staple single chain and biotin-modified staple single chain) are self-assembled. Mixing them allows them to self-assemble into DNA origami. The method of self-assembly of scaffold chains and staple single strands to form DNA origami is prior art. It is generally better to mix the scaffolding chai...

Embodiment 1

[0065] Example 1 Modified biotin without dT

[0066] Drop 2 μL of DNA origami modified with biotin (dT-free) (such as figure 1 a) On freshly dissociated mica, after 5 minutes of adsorption, in 50 μL 1×TAE / Mg 2+ In the buffer solution, use AFM’s liquid phase tap mode for imaging. After the imaging is stable, slowly inject 50 μL of 7.6nM streptavidin solution into the liquid tank, and continuously scan to observe the streptavidin and biotin on the origami site binding process, the observations are recorded in figure 1 b. Among them, the atomic force microscope is a NanoScope IIIa system (Digital Intrument, the United States), the scanning head of the atomic force microscope is a J-type scanning head, and the AFM probe is a NP-S silicon nitride tip (elasticity coefficient 0.58Nm -1 , Veeco Company), the imaging conditions were temperature 25°C, humidity 40%-50%, imaging parameters were scanning rate 2Hz, driving amplitude 400mV, voltage 0.3V, number of scanning lines 256, and ...

Embodiment 2

[0068] Embodiment 2 Modified biotin, containing multiple dT

[0069] The experimental procedure is the same as in Example 1, and the origami design pattern is as figure 2 a, the design site is the same as that in Example 1, except that it contains 5 dTs. When there are 5 dTs, there is only one case of biotin and avidin reaction on origami ( figure 2 b). Before the reaction, the origami had the same appearance, the length, width and height were 100nm, 70nm and 2.5nm, respectively. The surface is flat. After adding avidin, changes appeared on the origami surface over time, and after 30 minutes, almost all of the biotin-modified sites appeared circular bumps. The height is 2.8-3.3nm, which is the bound avidin molecule. from figure 2 The avidin binding site pattern on b can clearly determine the orientation of the origami on the mica surface. figure 2 b(1) is the image obtained after adding streptavidin and reacting for 5 minutes, figure 2 b(2) is the image obtained a...

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Abstract

The invention discloses a method related to DNA folded paper and a structure and an application thereof. The method comprises the following steps of: (1) preparing a DNA folded paper staple chain of a tail end modification reactant I, and arranging a connector between the tail end of the staple chain and the reactant I; (2) performing self-assembly on the staple chain obtained in the step (1) and a scaffold chain; (3) adding a reactant II which is specifically combined with the reactant I; and (4) detecting information on the combination of the reactant I and the reactant II. The connector is arranged between the reactant I and a staple short chain, so that the reaction locus of the reactant I which is effective to the reactant II on the DNA folded paper can be accurately controlled in particular to the DNA folded paper absorbed on a solid-liquid interface. The method can be taken as a detection method for a gene chip, and be used for specifically distributing reactants. The method can be used for detecting monomolecular reactions, and has extremely high sensitivity and specificity.

Description

technical field [0001] The invention belongs to the field of detection methods, in particular to a method involving DNA origami and its structure and application. Background technique [0002] Since Rothemund invented origami in 2006, it has aroused the interest of scientists in many fields including chemistry, biology, physics and material science, and has become one of the research hotspots in the field of nanometers. [0003] DNA origami has very excellent characteristics. In addition to the construction of delicate and addressable nanometer graphics through molecular design, many research groups have noticed another characteristic of origami - chirality. There is no difference in chirality in origami in solution, but when origami is adsorbed on the interface, there are differences in the orientation of the front and back sides, and they are mirror images of each other. The data obtained by different research groups are not the same. The experimental data of Rothemund a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/56C12M1/34G01N33/82G01N33/68G01N33/53
Inventor 吴娜李宾胡钧
Owner SHANGHAI INST OF APPLIED PHYSICS - CHINESE ACAD OF SCI
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