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

dc.contributor.author Jiang, Yong
dc.contributor.author Rabbi, Mahir
dc.contributor.author Mieczkowski, Piotr A
dc.contributor.author Marszalek, Piotr E
dc.coverage.spatial United States
dc.date.accessioned 2011-06-21T17:26:40Z
dc.date.issued 2010-09-23
dc.identifier http://www.ncbi.nlm.nih.gov/pubmed/20799746
dc.identifier.uri https://hdl.handle.net/10161/4074
dc.description.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.
dc.language eng
dc.language.iso en_US
dc.publisher American Chemical Society (ACS)
dc.relation.ispartof J Phys Chem B
dc.relation.isversionof 10.1021/jp105603k
dc.subject Calibration
dc.subject DNA
dc.subject DNA, Superhelical
dc.subject Electrophoresis, Agar Gel
dc.subject Microscopy, Atomic Force
dc.subject Plasmids
dc.title Separating DNA with different topologies by atomic force microscopy in comparison with gel electrophoresis.
dc.title.alternative
dc.type Journal article
duke.contributor.id Marszalek, Piotr E|0297721
dc.description.version Version of Record
duke.date.pubdate 2010-9-23
duke.description.issue 37
duke.description.volume 114
dc.relation.journal Journal of Physical Chemistry B
pubs.author-url http://www.ncbi.nlm.nih.gov/pubmed/20799746
pubs.begin-page 12162
pubs.end-page 12165
pubs.issue 37
pubs.organisational-group Duke
pubs.organisational-group Mechanical Engineering and Materials Science
pubs.organisational-group Pratt School of Engineering
pubs.publication-status Published
pubs.volume 114
dc.identifier.eissn 1520-5207


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