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

Stabile DNA-Motive, 1D und 2D Nanostrukturen aus kleinen kreisförmigen DNA-Moleküle gebaut

JoVE Journal
Chemistry

A subscription to JoVE is required to view this content. Sign in or start your free trial.

JoVE Journal Chemistry

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

Stabile DNA-Motive, 1D und 2D Nanostrukturen aus kleinen kreisförmigen DNA-Moleküle gebaut

Full Text

7,212 Views

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

Please note that some of the translations on this page are AI generated. Click here for the English version.

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.

Dieser Artikel stellt ein detailliertes Protokoll für T4 Ligatur und Geruchsstoffen Seite Reinigung der kleine kreisförmige DNA-Moleküle, Glühen und native Seitenanalyse der kreisförmigen Platten, Montage und AFM Imaging von 1D und 2D DNA Nanostrukturen sowie Agarose-gel Elektrophorese und Zentrifugation Reinigung des endlichen DNA Nanostrukturen.

Diese Methode erweitert die DNA-Nanotechnologie aus den Ausgangsmaterialien des linearen Oligonukleotids DAS in einige kreisförmige DAS. Die Hauptvorteile dieser technischen sind, dass es eine einfache, robuste und erschwinglich für die meisten Labore ist. Zur kreisförmigen DNA-Reinigung 20 Mikroliter Formamid zu einer Lösung frisch zubereiteter, intakter kreisförmiger DNA zu einem Endvolumen von 140 Mikrolitern mit Mischen hinzufügen.

Laden Sie 16 bis 20 Mikroliter kreisförmige DNA-Lösung auf jeweils sieben bis neun denaturierende PAGE-Gelbrunnen. Mischen Sie zwei Mikroliter Ladefarbstoff mit acht Mikrolitern der entsprechenden Vorläufer-Linear-DNA und injizieren Sie das Gemisch in einen separaten Referenzbrunnen. Dann laufen Sie das Gel für etwa zwei Stunden bei 10 Volt pro Zentimeter, stoppen Sie den Lauf, wenn das Xylol Cyanol FF etwa zwei Drittel des Gels beobachtet werden kann.

View the full transcript and gain access to thousands of scientific videos