Detection of DNA curvature by transverse pore gradient gel electrophoresis on PhastSystem gels

Zsuzsanna Buzás, Zsolt Boldogkol

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

It has been known since 1990 that DNA curvature can be recognized on transverse pore gradient gels by an intersection of 'Ferguson curves' with those of DNA size standards. The miniaturized PhastSystem polyacrylamide gels allow one to detect DNA curvature effortlessly and fast and at great economy of sample relative to alternative methods of electrophoresis. Using the transverse gradient gel electrophoresis method, it was found that the 660 bp length subfragment of the matrix attachment region (MAR) sequence of the chicken lysosyme gene migrates as a fragment of 800900 bp length. When subjected to digestion with the restriction enzyme HaeIII, the fragment gives rise to two species of 248 and 412 bp length, respectively. The Ferguson curves of both species intersect with those of DNA size standards, indicating that both exhibit curvature. Only the curvature of the 412 bp fragment conforms to prediction. Ethidium bromide abolishes the effect of curvature on the fragment, reducing its apparent size from 900 to 660, the value obtained by agarose gel electrophoresis.

Original languageEnglish
Pages (from-to)2838-2840
Number of pages3
JournalELECTROPHORESIS
Volume20
Issue number14
DOIs
Publication statusPublished - Nov 17 1999

Keywords

  • DNA curvature
  • PhastSystem

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry

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