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

Motivi di DNA stabile, 1D e 2D nanostrutture costruito da molecole di DNA circolare piccola

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Chemistry

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JoVE Journal Chemistry

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

Motivi di DNA stabile, 1D e 2D nanostrutture costruito da molecole di DNA circolare piccola

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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.

Questo articolo presenta un protocollo dettagliato per la legatura di T4 e denaturazione pagina purificazione di piccole molecole di DNA circolare, ricottura e nativo pagina analisi circolare piastrelle, montaggio e formazione immagine AFM di nanostrutture di DNA 1D e 2D, nonché dell'agarosi gel purificazione di elettroforesi e centrifugazione di nanostrutture di DNA finiti.

Questo metodo estende la nanotecnologia del DNA dai materiali sorgente dell'oligonucleotide lineare, DAS, in qualche DAS più circolare. I principali vantaggi di questi tecnici sono che è un semplice, robusto e conveniente per la maggior parte dei laboratori. Per la purificazione circolare del DNA, aggiungere 20 microlitri di formamide a una soluzione di DNA circolare appena preparato e intatto, ad un volume finale di 140 microlitri con miscelazione.

Caricare da 16 a 20 microlitri di soluzione circolare di DNA in ciascuno dei sette o nove pozzi di gel PAGE denaturanti. Mescolare due microlitri di colorante di carico con otto microlitri del corrispondente DNA lineare precursore e iniettare la miscela in un pozzo di riferimento separato. Quindi far funzionare il gel per circa due ore a 10 volt per centimetro, fermando la corsa quando lo xilene cianolo FF può essere osservato circa due terzi lungo il gel.

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