FAQ: Can I achieve digestion of a plasmid with a single SfiI site?

Trans reactions are aided by increasing DNA concentration (so you may achieve digestion by increasing the DNA concentration, but it may not be efficient) but, perversely, SfiI reaction can be hindered by increasing enzyme concentration. In reactions with enzyme in excess of DNA sites, each site becomes loaded with a SfiI tetramer, thus preventing the protein from binding the two sites necessary for full activity.

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