Separating DNA with different topologies by atomic force microscopy in comparison with gel electrophoresis.

Loading...
Thumbnail Image

Date

2010-09-23

Journal Title

Journal ISSN

Volume Title

Repository Usage Stats

268
views
343
downloads

Citation Stats

Abstract

Atomic force microscopy, which is normally used for DNA imaging to gain qualitative results, can also be used for quantitative DNA research, at a single-molecular level. Here, we evaluate the performance of AFM imaging specifically for quantifying supercoiled and relaxed plasmid DNA fractions within a mixture, and compare the results with the bulk material analysis method, gel electrophoresis. The advantages and shortcomings of both methods are discussed in detail. Gel electrophoresis is a quick and well-established quantification method. However, it requires a large amount of DNA, and needs to be carefully calibrated for even slightly different experimental conditions for accurate quantification. AFM imaging is accurate, in that single DNA molecules in different conformations can be seen and counted. When used carefully with necessary correction, both methods provide consistent results. Thus, AFM imaging can be used for DNA quantification, as an alternative to gel electrophoresis.

Department

Description

Provenance

Citation

Published Version (Please cite this version)

10.1021/jp105603k

Publication Info

Jiang, Yong, Mahir Rabbi, Piotr A Mieczkowski and Piotr E Marszalek (2010). Separating DNA with different topologies by atomic force microscopy in comparison with gel electrophoresis. J Phys Chem B, 114(37). pp. 12162–12165. 10.1021/jp105603k Retrieved from https://hdl.handle.net/10161/4074.

This is constructed from limited available data and may be imprecise. To cite this article, please review & use the official citation provided by the journal.


Unless otherwise indicated, scholarly articles published by Duke faculty members are made available here with a CC-BY-NC (Creative Commons Attribution Non-Commercial) license, as enabled by the Duke Open Access Policy. If you wish to use the materials in ways not already permitted under CC-BY-NC, please consult the copyright owner. Other materials are made available here through the author’s grant of a non-exclusive license to make their work openly accessible.