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. 2019 May 10;58(23):2675–2681. doi: 10.1021/acs.biochem.9b00063

Figure 3.

Figure 3

Nb.BssSI background restriction activity can be suppressed by adding a PTO bond to the template strand. (a) Nickase Nb.BssSI is supposed to cut at the site indicated in red. The restriction enzyme BssSI additionally cuts at the site indicated in blue. We tested four templates, T1u (purple), T1 (blue), T1 (green), and T1fp (yellow), with increasing number of PTOs around the BssSI site. (b) EvaGreen fluorescence versus time for rEXPAR experiments with templates in panel a. Conditions: 8 U/mL pol, 200 U/mL nick, 100 nM exo, and 0.4 mM dNTPs.