A rapid high-resolution method for resolving DNA topoisomers
Abstract Objective Agarose gel electrophoresis has been the mainstay technique for the analysis of DNA samples of moderate size. In addition to separating linear DNA molecules, it can also resolve different topological forms of plasmid DNAs, an application useful for the analysis of the reactions of...
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doaj-fb45c1ac20f040ae8c3d392c376015152020-11-25T02:37:14ZengBMCBMC Research Notes1756-05002018-01-011111710.1186/s13104-018-3147-6A rapid high-resolution method for resolving DNA topoisomersLesley A. Mitchenall0Rachel E. Hipkin1Michael M. Piperakis2Nicolas P. Burton3Anthony Maxwell4Department of Biological Chemistry, John Innes CentreQiagen Ltd., Skelton HouseDepartment of Biological Chemistry, John Innes CentreInspiralis Ltd, Innovation CentreDepartment of Biological Chemistry, John Innes CentreAbstract Objective Agarose gel electrophoresis has been the mainstay technique for the analysis of DNA samples of moderate size. In addition to separating linear DNA molecules, it can also resolve different topological forms of plasmid DNAs, an application useful for the analysis of the reactions of DNA topoisomerases. However, gel electrophoresis is an intrinsically low-throughput technique and suffers from other potential disadvantages. We describe the application of the QIAxcel Advanced System, a high-throughput capillary electrophoresis system, to separate DNA topoisomers, and compare this technique with gel electrophoresis. Results We prepared a range of topoisomers of plasmids pBR322 and pUC19, and a 339 bp DNA minicircle, and compared their separation by gel electrophoresis and the QIAxcel System. We found superior resolution with the QIAxcel System, and that quantitative analysis of topoisomer distributions was straightforward. We show that the QIAxcel system has advantages in terms of speed, resolution and cost, and can be applied to DNA circles of various sizes. It can readily be adapted for use in compound screening against topoisomerase targets.http://link.springer.com/article/10.1186/s13104-018-3147-6Capillary electrophoresisDNA topoisomerasesPlasmidsMinicircles |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Lesley A. Mitchenall Rachel E. Hipkin Michael M. Piperakis Nicolas P. Burton Anthony Maxwell |
spellingShingle |
Lesley A. Mitchenall Rachel E. Hipkin Michael M. Piperakis Nicolas P. Burton Anthony Maxwell A rapid high-resolution method for resolving DNA topoisomers BMC Research Notes Capillary electrophoresis DNA topoisomerases Plasmids Minicircles |
author_facet |
Lesley A. Mitchenall Rachel E. Hipkin Michael M. Piperakis Nicolas P. Burton Anthony Maxwell |
author_sort |
Lesley A. Mitchenall |
title |
A rapid high-resolution method for resolving DNA topoisomers |
title_short |
A rapid high-resolution method for resolving DNA topoisomers |
title_full |
A rapid high-resolution method for resolving DNA topoisomers |
title_fullStr |
A rapid high-resolution method for resolving DNA topoisomers |
title_full_unstemmed |
A rapid high-resolution method for resolving DNA topoisomers |
title_sort |
rapid high-resolution method for resolving dna topoisomers |
publisher |
BMC |
series |
BMC Research Notes |
issn |
1756-0500 |
publishDate |
2018-01-01 |
description |
Abstract Objective Agarose gel electrophoresis has been the mainstay technique for the analysis of DNA samples of moderate size. In addition to separating linear DNA molecules, it can also resolve different topological forms of plasmid DNAs, an application useful for the analysis of the reactions of DNA topoisomerases. However, gel electrophoresis is an intrinsically low-throughput technique and suffers from other potential disadvantages. We describe the application of the QIAxcel Advanced System, a high-throughput capillary electrophoresis system, to separate DNA topoisomers, and compare this technique with gel electrophoresis. Results We prepared a range of topoisomers of plasmids pBR322 and pUC19, and a 339 bp DNA minicircle, and compared their separation by gel electrophoresis and the QIAxcel System. We found superior resolution with the QIAxcel System, and that quantitative analysis of topoisomer distributions was straightforward. We show that the QIAxcel system has advantages in terms of speed, resolution and cost, and can be applied to DNA circles of various sizes. It can readily be adapted for use in compound screening against topoisomerase targets. |
topic |
Capillary electrophoresis DNA topoisomerases Plasmids Minicircles |
url |
http://link.springer.com/article/10.1186/s13104-018-3147-6 |
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