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. 2015 Feb 19;43(6):3272–3285. doi: 10.1093/nar/gkv078

Figure 1.

Figure 1.

A real-time fluorescence assay for monitoring open-complex formation. (A) The assay utilizes a Cy3 dye at the +2 position of the non-template strand of a rrnAP3 ribosomal RNA promoter DNA template. The Cy3 has a baseline fluorescence in the free and closed complexes that is enhanced upon the formation of open complex. (B) EcoRNAP-dependent (21 nM) fluorescence enhancement in the presence (green) and absence (red) promoter sequence at 25°C. (C) EcoRNAP-dependent (21 nM) fluorescence enhancement at 10°C (blue), 25°C (green) and 37°C (red). (D) MboRNAP-dependent (100 nM) fluorescence enhancement in the presence (green) and absence (red) of promoter sequence at 25°C. (E) MboRNAP-dependent (100 nM) fluorescence enhancement at 10°C (blue), 25°C (green) and 37°C (red).