Table 1.

Cas9 cleavage of torsionally relaxed and negatively supercoiled DNA templates

	Linear	Nicked	Negatively supercoiled
	Cleavage	Cleavage	Cleavage	Nicking
λ1	0.91 ± 0.01	0.92 ± 0.01	1.00	n.d.
13-20mm	n.d.	n.d.	0.35 ± 0.04	0.34 ± 0.01
18CA	0.88 ± 0.01	0.90 ± 0.02	1.00	n.d.
3CA	0.97 ± 0.01	0.94 ± 0.03	1.00	n.d.
8TA	0.98 ± 0.01	1.00	1.00	n.d.
1-4mm	n.d.	n.d.	0.02 ± 0.01	0.19 ± 0.05

Linear and nicked reaction templates were generated by treating the negatively supercoiled plasmid DNA with endonucleases DraIII and Nb.BsrDI, respectively. Fractional cleavage is given as mean ± SD for n = 3 replicates. The detection limit of the assay is ∼1% cleavage or nicking; SD is not given when all three measurements agree within <1%. n.d., not detected.