Table 1.

Properties of DNA-guided TtAgo binding to DNA targets with different extents of complementarity to the guide-strand at various temperatures

	kon (M−1 s−1)
	23°C	37°C	45°C	55°C
Fully complementary	6.5 ± 0.1 × 107	8.5 ± 0.1 × 107	12.0 ± 0.1 × 107	11.1 ± 0.1 × 107
Seed-matched	7.0 ± 0.1 × 107	6.9 ± 0.1 × 107	9.0 ± 0.1 × 107	12.0 ± 0.1 × 107
Seed-matched + 3′ sup. pairing	7.5 ± 0.1 × 107	5.5 ± 0.1 × 107	9.0 ± 0.1 × 107	13.5 ± 0.1 × 107

	koff (s−1)
	23°C	37°C	45°C	55°C
Fully complementary	« PB	« PB	« PB	« PB
Seed-matched	0.10 ± 0.01	0.61 ± 0.01	2.0 ± 0.1	18.0 ± 0.3
Seed-matched + 3′ sup. pairing	0.09 ± 0.01	0.67 ± 0.01	2.0 ± 0.1	13.2 ± 0.3

Values of k_on and k_off were derived from data collected from several hundred individual DNA target molecules ( > 1100); standard error from bootstrapping is reported. Representative fluorescence intensity time traces and rastergrams summarizing traces of individual target molecules are shown in Supplementary Fig. 2