Separation of plasmid DNA isoforms using centrifugal ultrafiltration

Ehsan Espah Borujeni, Andrew Zydney

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Centrifugal ultrafiltration is a well-established method for concentrating and purifying DNA. Here, we describe the use of centrifugal ultrafiltration for the separation of plasmid DNA isoforms based on differences in elongational flexibility of the supercoiled, open-circular, and linear plasmids. Transmission of each isoform is minimal below a critical value of the filtration velocity, which is directly related to the magnitude of the centrifugal speed and the system geometry. A discontinuous diafiltration process was used to enrich the desired isoform, as determined by agarose gel electrophoresis. The simplicity and efficacy of this membrane-based separation are attractive for multiple applications requiring the use of separated DNA isoforms.

Original languageEnglish (US)
Pages (from-to)49-56
Number of pages8
JournalBioTechniques
Volume53
Issue number1
DOIs
StatePublished - Jul 1 2012

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Ultrafiltration
Protein Isoforms
Plasmids
DNA
Agar Gel Electrophoresis
Electrophoresis
Sepharose
Gels
Membranes
Geometry

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Borujeni, Ehsan Espah ; Zydney, Andrew. / Separation of plasmid DNA isoforms using centrifugal ultrafiltration. In: BioTechniques. 2012 ; Vol. 53, No. 1. pp. 49-56.
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Separation of plasmid DNA isoforms using centrifugal ultrafiltration. / Borujeni, Ehsan Espah; Zydney, Andrew.

In: BioTechniques, Vol. 53, No. 1, 01.07.2012, p. 49-56.

Research output: Contribution to journalArticle

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