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. 2013 Jul 1;41(17):8182–8195. doi: 10.1093/nar/gkt588

Figure 6.

Figure 6.

Binding and thermostabilization of a dsDNA fragment and an RNA:DNA hybrid by Sso7d. (A) Binding of Sso7d to a dsDNA fragment and an RNA:DNA hybrid. Sso7d was incubated for 10 min at 25°C with a radiolabeled 30-bp dsDNA fragment or a 30-bp RNA:DNA hybrid (2 nM). Samples were loaded onto a 5% polyacrylamide gel and electrophoresed in 0.1× TBE. Gels were dried and exposed to radiographic film. Sso7d concentrations were 0, 0.05, 0.1, 0.5, 1, 5, 10, 50 and 90 μM, respectively. (B) A plot of the Sso7d-bound fraction of the radiolabeled probe versus input Sso7d concentration. (C and D) Effect of Sso7d on the thermal stability of a dsDNA fragment and an RNA:DNA hybrid. Thermal denaturation of a 15-bp dsDNA fragment or an RNA:DNA hybrid in the presence of various amounts of Sso7d was determined by monitoring changes in UV absorbance at 260 nm.