TY - JOUR
T1 - Quantitation of ethidium-stained closed circular DNA in agarose gels
AU - Shubsda, Michael F.
AU - Goodisman, Jerry
AU - Dabrowiak, James C.
PY - 1997/2/1
Y1 - 1997/2/1
N2 - 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 ~ 20% more than form III DNA.
AB - 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 ~ 20% more than form III DNA.
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U2 - 10.1016/S0165-022X(96)01204-3
DO - 10.1016/S0165-022X(96)01204-3
M3 - Article
C2 - 9089386
AN - SCOPUS:0031052217
SN - 0165-022X
VL - 34
SP - 73
EP - 79
JO - Journal of Biochemical and Biophysical Methods
JF - Journal of Biochemical and Biophysical Methods
IS - 1
ER -