Unidirectional pulsed-field electrophoresis of single- and double-stranded DNA in agarose gels: Analytical expressions relating mobility and molecular length and their application in the measurement of strand breaks

John Clark Sutherland, Denise C. Monteleone, Jo Ann H. Mugavero, John Trunk

Research output: Contribution to journalArticle

54 Scopus citations

Abstract

Unidirectional pulsed-field electrophoresis improves the separation of single-stranded DNA molecules longer than 20 kilobases (kb) in alkaline agarose gels compared to static-field electrophoresis. The greatest improvement in separation is for molecules longer than 100 kb. The improved resolution of long molecules with unidirectional pulsed-field electrophoresis makes possible the measurement of lower frequencies of single-strand breaks. The analytical function that relates the length and mobility of single-stranded DNA electrophoresed with a static field also applies to unidirectional pulsed field separations. Thus, the computer programs used to measure single-strand breaks are applicable to both undirectional pulsed- and static-field separations. Unidirectional pulsed-field electrophoresis also improves the separation of double-stranded DNA in neutral agarose gels. The function relating molecular length and mobility for double-stranded DNA separated by unidirectional pulsed-field electrophoresis is a superset of the function for single-stranded DNA. The coefficients of this function can be determined by iterative procedures.

Original languageEnglish (US)
Pages (from-to)511-520
Number of pages10
JournalAnalytical Biochemistry
Volume162
Issue number2
DOIs
StatePublished - May 1 1987
Externally publishedYes

Keywords

  • DNA damage
  • DNA mobility
  • agarose gel electrophoresis
  • pulsed-field electrophoresis
  • single-strand breaks

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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