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Prenicking of the 12-substrate eliminates its trans effect on the hairpin formation of the partner 23-substrate in trans. The left panel shows the cleavage of the 23-high paired with a prenicked 12-high over a period of 30 min (lanes 1 through 5). The right panel shows the cleavage of the same 23-high paired with a prenicked 12-low over the same period of time (lanes 6 through 10). All other symbols are as in previous figures.
