Table 2.

Steady State kinetics incorporation of 2APTP and 4CIN-TP by Kf⁻ opposite the four natural DNA bases.

Nucleotide/Template base	KM (μM)	Vmax (nM min−1)	kcat (min−1)	Catalytic efficiency (kcat/KM) (μM−1 min−1)
2AP-TP
vs TempA	175.3 ± 27.4	0.0127 ± 0.0060	0.0127	7.23×10−5
vs TempC	195.3 ± 10.8	0.182 ± 0.053	0.182	9.44×10−4
vs TempG	98.3 ± 63.8	0.0167 ± 0.008	0.0167	1.82×10−4
vs TempT	3.67 ± 0.54	1.16 ± 0.59	1.16	3.19×10−1
4CIN-TP
vs TempA	42.1 ± 12.6	0.0952 ± 0.0391	0.0952	2.49×10−3
vs TempC	34.6 ± 8.9	0.0321 ± 0.0238	0.0321	9.47×10−4
vs TempG	40.7 ± 9.7	0.0185 ± 0.0151	0.0185	4.33×10−4
vs TempT	78.7 ± 15.4	0.0858 ± 0.0051	0.0858	1.11×10−3

These kinetics were done in triplicate at 25 °C with 1 nM Kf⁻, and 100 nM Cy5-labelled primer/template.