Stable DNA Motifs, 1D and 2D Nanostructures Constructed from Small Circular DNA Molecules

Xin Guo; Xue-Mei Wang; Shou-Jun Xiao

doi:10.3791/58744

Motivos de ADN estable, 1D y 2D nanoestructuras construyen a partir de moléculas de ADN Circular ...

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JoVE Journal Chemistry

Stable DNA Motifs, 1D and 2D Nanostructures Constructed from Small Circular DNA Molecules

Motivos de ADN estable, 1D y 2D nanoestructuras construyen a partir de moléculas de ADN Circular pequeño

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09:32 min

April 12, 2019

DOI: 10.3791/58744-v

Xin Guo¹, Xue-Mei Wang¹, Shou-Jun Xiao¹

¹State Key Laboratory of Coordination Chemistry, School of Chemistry and Chemical Engineering,Nanjing University

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Overview

This article presents a detailed protocol for T4 ligation and purification of small circular DNA molecules, as well as the analysis and imaging of DNA nanostructures. The methods described are simple, robust, and affordable for most laboratories.

Key Study Components

Area of Science

DNA nanotechnology
Circular DNA purification
Electrophoresis techniques

Background

Extends DNA nanotechnology from linear to circular DNA.
Focuses on purification and analysis techniques.
Utilizes denaturing PAGE for DNA separation.
Offers insights into assembling DNA nanostructures.

Purpose of Study

To provide a protocol for circular DNA purification.
To demonstrate methods for analyzing DNA nanostructures.
To enhance accessibility of DNA nanotechnology techniques.

Methods Used

T4 ligation for DNA assembly.
Denaturing PAGE for purification of circular DNA.
AFM imaging for structural analysis of DNA nanostructures.
Agarose gel electrophoresis for further purification.

Main Results

Successful purification of circular DNA using formamide.
Effective separation of DNA samples via denaturing PAGE.
Visualization of 1D and 2D DNA nanostructures.
Demonstrated robustness and simplicity of the methods.

Conclusions

The protocols are accessible for most laboratories.
Methods can be applied to various DNA nanotechnology projects.
Enhances understanding of circular DNA applications.

Frequently Asked Questions

What is T4 ligation?

T4 ligation is a method used to join two DNA fragments together using T4 DNA ligase.

How is circular DNA purified?

Circular DNA is purified by adding formamide to the solution and using denaturing PAGE for separation.

What is denaturing PAGE?

Denaturing PAGE is a technique used to separate nucleic acids based on size under denaturing conditions.

What are the advantages of using circular DNA?

Circular DNA can be more stable and efficient for certain applications in DNA nanotechnology.

What is AFM imaging?

AFM imaging is a technique that allows for high-resolution visualization of nanoscale structures, including DNA nanostructures.

Este artículo presenta un protocolo detallado para ligadura de T4 y desnaturalización página purificación de moléculas pequeñas de ADN circular, recocido y del análisis de la página nativa de Tejas circular, montaje y proyección de imagen de AFM de nanoestructuras de ADN de 1D y 2D, así como de agarosa gel purificación de electroforesis y centrifugación de nanoestructuras de ADN finito.

Este método extiende la nanotecnología del ADN a partir de los materiales de origen del oligonucleótido lineal, DAS, a un DAS más circular. Las principales ventajas de estos técnicos son que es simple, robusto y asequible para la mayoría de los laboratorios. Para la purificación circular del ADN, añada 20 microlitros de formamida a una solución de ADN circular recién preparado e intacto, a un volumen final de 140 microlitros con mezcla.

Cargue de 16 a 20 microlitros de solución circular de ADN a cada uno de siete a nueve pozos de gel PAGE desnaturalizadores. Mezclar dos microlitros de tinte de carga con ocho microlitros del ADN lineal precursor correspondiente, e inyectar la mezcla en un pozo de referencia separado. A continuación, ejecute el gel durante unas dos horas a 10 voltios por centímetro, deteniendo la carrera cuando el xileno ciaol FF se puede observar alrededor de dos tercios por el gel.

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