Determination of plasmid copy number by fluorescence densitometry.
A simple and reliable method for the determination of plasmid copy numbers by direct fluorescence densitometry of ethidium bromide-stained electrophoretic gels was developed. In developing the method, the following parameters were evaluated and controlled: plasmid DNA trapping in the linear chromosomal DNA, staining-destaining kinetics for ethidium bromide, linearity of the fluorescence response, and the effect of the molecular topology of DNA on ethidium bromide binding to DNA in agarose.
D004269:DNA, Bacterial / Q000235:genetics; D003720:Densitometry / Q000379:methods*; D004587:Electrophoresis, Agar Gel; D004996:Ethidium; D005453:Fluorescence; D005784:Gene Amplification; D005821:Genetic Techniques; D009690:Nucleic Acid Conformation; D010957:Plasmids; D013211:Staphylococcus aureus / Q000235:genetics
D004269:DNA, Bacterial; D004996:Ethidium