Table 3.

Steady-state kinetics of single nucleotide extension past 2′-F dG:dC, 2′-F dG:dT and N⁷-CH₃ 2′-F dG:dC, and N⁷-CH₃ 2′-F dG:dT base pairs by hpol η

The oligonucleotides used were as follows,

5′-FAM-CGGGCTCGTAAGCGTCATC-3′ 3′-GCCCGAGCATTCGCAGTAXTACT-5′

5′-FAM-CGGGCTCGTAAGCGTCATT-3′ 3′-GCCCGAGCATTCGCAGTAXTACT-5′

where X represents 2′-F dG and N⁷-CH₃ 2′-F dG.

Template base	Pairing	dNTP	kcat	Km	kcat/Km
			min−1	μm	μm−1 min−1
2′-F dG	Mispair	dATP	1.1 ± 0.1	4.2 ± 1.2	0.26 ± 0.08
N7-CH3 2′-F dG		dATP	0.49 ± 0.04	7.1 ± 2.4	0.07 ± 0.02
2′-F dG	Correct	dATP	1.1 ± 0.1	15 ± 3	0.07 ± 0.02
N7-CH3 2′-F dG		dATP	2.1 ± 0.1	1.4 ± 0.3	1.5 ± 0.3
2′-F dG	Correct	dCTP	0.95 ± 0.06	5.5 ± 2.2	0.17 ± 0.07
N7-CH3 2′-F dG		dCTP	0.75 ± 0.03	4.6 ± 1.1	0.16 ± 0.04
2′-F dG	Correct	dGTP	2.1 ± 0.06	6.9 ± 1.0	0.3 ± 0.1
N7-CH3 2′-F dG		dGTP	0.97 ± 0.03	3.8 ± 0.7	0.26 ± 0.1
2′-F dG	Correct	dTTP	1.0 ± 0.1	3.4 ± 0.7	0.3 ± 0.10
N7-CH3 2′-F dG		dTTP	1.6 ± 0.1	0.96 ± 0.47	1.7 ± 0.8