Figure 2 Time-dependent digestion of plasmids with two PaqCI target sites indicate a sequential cleavage profile

The reaction contained 1 ug substrate DNA and 2 units/ug PaqCI endonuclease in rCutSmart buffer at 37C. Digest time points were quenched at 0.25 min, 0.5 min, 1 min, 3 min, 5 min, 10 min, 30 min, and 60 minutes with 1 ug dual site substrate and 2 units/ug PaqCI as indicated above each lane. The mobilities of the super coil (SC) and the cleaved linear (Lin, Lin1 and Lin2) forms of the plasmid are indicated in the middle. Below the gels is a cartoon of the substrate and reaction products shown in the gel.