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  • Title: Quantitation of ethidium-stained closed circular DNA in agarose gels.
    Author: Shubsda MF, Goodisman J, Dabrowiak JC.
    Journal: J Biochem Biophys Methods; 1997 Feb 01; 34(1):73-9. PubMed ID: 9089386.
    Abstract:
    The fluorescence of ethidium bromide (EB) bound to equimolar amounts of supercoiled form I and unstrained linear form III pBR322, SV40 and PM2 DNA in agarose gels has been measured by scanning a photographic negative of the gel with a microdensitometer. For SV40 and PM2 DNA, commonly used staining conditions cause both forms, i.e. linear and supercoiled, to fluoresce to the same extent. This obviates the need to use a correction factor for the fluorescence of form I DNA when measuring the amount of this form relative to the amounts of unstrained forms in agarose gels. In the case of PBR322 DNA, form I was found to fluoresce approximately 20% more than form III DNA.
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