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. 2016 Jul 1;7(7):32. doi: 10.3390/genes7070032

Figure 1.

Figure 1

The effect of etoposide concentration on Top2cc trapping. The reactions (5 μL) contain: pGT2 plasmid DNA (10 ng/μL), Top2 (1 unit/μL; USB, OH) or buffer and the indicated concentrations of etoposide (ETP) (Sigma, St. Louis, MO, USA) (the Top2 reactions also contained 1 mM ATP). After incubation at 37 °C for 10 min, the reactions were terminated with 1.5 μL of 4% SDS + 50 mM EDTA and then treated with 7.5 μL proteinase K (1 μg/μL; Sigma, St. Louis, MO, USA) at room temperature for 2 h. The products were separated on a 1% TAE-agarose gel at 80 volts for 3 h and detected by SYBR Gold staining (Invitrogen, Carlsbad, CA, USA).