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. Author manuscript; available in PMC: 2016 Jan 1.
Published in final edited form as: Plasmid. 2014 Nov 5;0:7–16. doi: 10.1016/j.plasmid.2014.11.001

Figure 5. Single-stranded DNA staining and the effect of replication origins on plasmid DNA.

Figure 5

A) Left panel: ethidium bromide-stained DNA by pre-incorporation of the dye into the gel. Lane 1, double-stranded M13; lane 2, single-stranded M13 DNA; lane 3, pColE3 miniprep; and lane 4, CsCl-purified pColE3. In lanes 2 & 3, arrows indicate the diffuse ssDNA bands running as a lower molecular weight with ethidium bromide pre-incorporated into the gel. Right panel: acridine orange-stained DNA. Lane 1, dsM13; lane 2, ssM13; lane 3, pColE3; and lane 4, CsCl-purified pColE3. Arrows indicate the diffuse bands associated with M13 ssDNA and pColE3 that are shifted to a higher apparent molecular weight in the absence of ethidium bromide. B) pUC19 constructs containing pColE3 components. C) Left panel: ethidium bromide-stained DNA by pre-incorporation of the dye into the gel. Lane 1, pUC19; lane 2, construct 1; lane 3, construct 2; lane 4, construct 3; lane 5, construct 4. In lanes 4, an arrow indicate one bands running as a slightly lower molecular weight with ethidium bromide incorporated into the gel during electrophoresis. Right panel: acridine orange-stained DNA. Lane 1, pUC19; Lane 2, construct 1; lane 3, construct 2; lane 4, construct 3; lane 5, construct 4. In lanes 4, an arrow indicate one bands running as a slightly higher apparent molecular weight in the absence ethidium bromide incorporated into the gel during electrophoresis. D) Relative concentrations of DNA produced by pUC19, pColE3, construct 1 and construct 3.