A rapid high-resolution method for resolving DNA topoisomers View Full Text


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Article Info

DATE

2018-01-16

AUTHORS

Lesley A. Mitchenall, Rachel E. Hipkin, Michael M. Piperakis, Nicolas P. Burton, Anthony Maxwell

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. More... »

PAGES

37

References to SciGraph publications

  • 2011-12-16. Interfacial inhibitors: targeting macromolecular complexes in NATURE REVIEWS DRUG DISCOVERY
  • 2011-11-23. All tangled up: how cells direct, manage and exploit topoisomerase function in NATURE REVIEWS MOLECULAR CELL BIOLOGY
  • 2011-09-09. Exploiting bacterial DNA gyrase as a drug target: current state and perspectives in APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
  • 1999-03-23. Overexpression and Purification of Bacterial DNA Gyrase in DNA TOPOISOMERASE PROTOCOLS
  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1186/s13104-018-3147-6

    DOI

    http://dx.doi.org/10.1186/s13104-018-3147-6

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1100433844

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/29338757


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