Electrophoretic behavior of plasmid DNA in the presence of various intercalating dyes

Journal of Chromatography. a
Fuquan DangYoshinobu Baba

Abstract

In the present study, the electrophoretic behavior of linear, supercoiled and nicked circular plasmid DNA in the presence of various intercalating dyes was characterized using pGL3 plasmid DNA as a model. The enzymatic digestion of pGL3 plasmid DNA with HindIIIwas monitored by capillary electrophoresis coupled with laser-induced fluorescence detection (CE-LIF). Nicked circular plasmid DNA was found to be relatively sensitive to enzymes, and was almost digested into the linear conformer after 10-min incubation, indicating that nicked circular plasmid DNA has little chance of targeting and entering the cell nucleus. Partly digested plasmid DNA containing only linear and supercoiled conformers can be used as a standard to confirm the migration order of plasmid DNA. In methylcellulose (MC) solution with YO-PRO-1 or YOYO-1, linear plasmid DNA eluted first, followed by supercoiled and nicked plasmid DNA, and nicked plasmid DNA eluted as a broad peak. With SYBR Green 1, nicked plasmid DNA eluted first as three sharp peaks, followed by linear and supercoiled plasmid DNA. The nuclear plasmid DNA from two transfected cell lines was successfully analyzed using the present procedure. Similar results were obtained with an analysis time of s...Continue Reading

References

Aug 9, 1996·Journal of Chromatography. B, Biomedical Applications·Z NackerdienD Atha
Jun 29, 2000·Journal of Chromatography. B, Biomedical Sciences and Applications·S A NevinsZ E Nackerdien

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Citations

Feb 19, 2013·Electrophoresis·Alberto ValdésAlejandro Cifuentes
Jun 1, 2011·Electrophoresis·Airong FengPing Zhou
Sep 28, 2021·Electrophoresis·Urh ČernigojAleš Štrancar

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