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. 2005 Mar 14;33(5):1532–1543. doi: 10.1093/nar/gki293

Figure 9.

Figure 9

Effect of the downstream template base N20 on chain extension reaction catalyzed by KF exo− starting from the unnatural base pair 7-Ni(in primer):T(in template). Experiments were investigated on different natural templates (100 nM) varying by the nature of the base N20 (N = T, sequence 13; N = A, sequence 20; N = C, sequence 21; N = G, sequence 22) hybridized to 18mer primer (sequence 12, 50 nM). Addition of one d(7-Ni) to the primer was carried out using KF exo− (0.0625 U/μl) in presence of d(7-Ni)TP (1 mM) for 5 min at 30°C (lanes 2, 6, 10 and 14). The second step of reaction was performed using KF exo− (0.0125 U/μl) in the presence of one single dNTP (lanes 3, 7, 11 and 15) or in the presence of the mixture of the four dNTPs (lanes 4, 8, 12 and 16) for 20 min at 30°C.