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Method for forming dimer structure by assembling DNA origami template and nanometer gold cube based on surface enhanced Raman effect

A surface-enhanced Raman and nano-gold technology, which is applied in the field of DNA nanometers, can solve the problems of assembly neglect and achieve the effects of rich diversity, precise space addressability, and efficient and precise assembly

Active Publication Date: 2019-07-26
NANJING UNIV OF POSTS & TELECOMM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, among various plasmonic metal particles, the assembly of non-spherical anisotropic nanoparticles with angularity has been relatively neglected.

Method used

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  • Method for forming dimer structure by assembling DNA origami template and nanometer gold cube based on surface enhanced Raman effect
  • Method for forming dimer structure by assembling DNA origami template and nanometer gold cube based on surface enhanced Raman effect
  • Method for forming dimer structure by assembling DNA origami template and nanometer gold cube based on surface enhanced Raman effect

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Preparation of triangular DNA origami with specific sites

[0032] Such as figure 1 Shown: Using phage M13mp18 single-stranded DNA as the backbone strand, it anneals with more than two hundred short scaffold strands and dozens of capture strands to form a triangular DNA origami. Its specific operation is:

[0033] (1) Mix M13mp18 bacteriophage circular single-stranded DNA, unmodified staple strand, and terminal-modified capture strand (CaptureDNA) at a molar ratio of 1:10:10, that is, the added volumes are 2.5 μL and 5 μL, respectively. μL, 5 μL, then 10 μL 10×TAE-Mg 2+ Buffer (Mg 2+ concentration 12.5 mol / L), add ultrapure water to a final volume of 100 μL, shake well.

[0034] (2) Place the mixed solution in step (1) in a PCR instrument and anneal at a rate of 0.1°C / 10 s from 95°C to 20°C. After the reaction, use a 100 kDa ultrafiltration tube to centrifuge to remove excess staple chains. 4 ℃ for later use.

[0035] Among them, the sequence of the cap...

Embodiment 2

[0039] Embodiment 2: Preparation of nano gold cubes modified by thiol DNA

[0040] In a certain buffer environment, thiol DNA single strands were added to modify the surface of the nano-gold cube, and the assembly was completed by adding salt (NaCl) aging to obtain a nano-gold cube covered with thiol DNA. Its specific operation is:

[0041] (1) Take 200 μL of 10 nm gold particles in a 1.5 mL centrifuge tube, centrifuge at 12000 rpm for 15 min, remove 120 μL of supernatant, add 10 μL of thiol DNA and 10 μL of 5×TBE (89 mM Tris, 89 mM boric acid, 2 mMEDTA, pH 8.0) buffer solution, vortexed to mix.

[0042] (2) Place the sample in step (1) in a mixer, and incubate at 37°C and 300 rpm for 4-6 hours.

[0043] (3) Add 10 μL of 3M NaCl solution to the sample incubated in step (2). Pay attention to adding 4 times, each adding 2.5 μL of 3 M NaCl solution at intervals of 30 minutes. Incubate overnight at 37°C. The purpose of adding salt (NaCl) step by step is to enable the assembly o...

Embodiment 3

[0045] Example 3: Preparation of three dimer configurations

[0046] Such as figure 1 As shown, the thiol DNA-modified nano gold cube prepared in Example 2 was mixed with the triangular DNA origami prepared in Example 1, annealed and assembled. Its specific operation is:

[0047] The DNA origami and nano-gold cubes modified with sulfhydryl DNA were uniformly mixed in 1×TAE-Mg at a molar ratio of 1:2. 2+ In the buffer, the sample was placed in a PCR machine and annealed for 11 h. The annealing program was to cool from 45 °C to 20 °C at a rate of -0.1 °C / 10 s, and cycled for 4 cycles to completely hybridize the DNA origami and the nano-gold cube. The capture chains at different positions were designed on the surface, and finally assembled to form three kinds of nano-gold cubic dimers: face-face (FTF), face-edge (FTS) and edge-edge (STS).

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Abstract

The invention discloses a method for forming a dimer structure by assembling a DNA origami template and a nanometer gold cube based on a surface enhanced Raman effect. The method specifically comprises the five steps of preparation of triangular DNA origami with specific sites; preparation of a sulfydryl DNA modified nanometer gold cube; preparation of three dimer configurations; and high efficiency precise assembly by using agarose gel electrophoresis and a transmission electron microscope representational structure, and surface enhanced Raman scattering by using a scanning electron microscope and dark-field microscope co-location detection assembly structure. The method solves the problems of low assembly repeatability and poor structural stability, a new idea is also provided for construction of nanometer optical materials due to establishment of the method, and the method is of great significance to research of nanophotonics.

Description

technical field [0001] The invention belongs to the field of DNA nanotechnology, and in particular relates to a method for assembling nano-gold cubes to form a dimer structure based on a surface-enhanced Raman effect-based DNA origami template. Background technique [0002] When some molecules are adsorbed on the surface of a specific substance (such as Au, Ag or Cu), the Raman spectrum signal intensity of the molecules will increase significantly. This Raman scattering enhancement phenomenon is called surface-enhanced Raman scattering effect. , in the past decade, surface-enhanced Raman scattering technology has been widely used in many fields such as surface adsorption, electrochemistry, catalytic reaction, biochemical sensor, biomedical detection, trace detection and analysis, etc. Strong light field enhancement and effective light collection effectively solve the problem of low sensitivity of Raman spectroscopy. However, this technique has the problems of poor reproduci...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/65C12P19/34
CPCG01N21/658C12P19/34
Inventor 汪联辉腾楠晁洁
Owner NANJING UNIV OF POSTS & TELECOMM
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