Projan S J, Carleton S, Novick R P
Plasmid. 1983 Mar;9(2):182-90. doi: 10.1016/0147-619x(83)90019-7.
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.
开发了一种通过对溴化乙锭染色的电泳凝胶进行直接荧光密度测定来确定质粒拷贝数的简单可靠方法。在开发该方法时,对以下参数进行了评估和控制:质粒DNA在线性染色体DNA中的截留、溴化乙锭的染色-脱色动力学、荧光响应的线性以及DNA分子拓扑结构对溴化乙锭与琼脂糖中DNA结合的影响。