Optimal activity is observed in ThermoPol reaction buffer (200% activity). The unit is assayed in a non-optimal buffer taken from the literature. The enzyme exhibits 50% activity in NEBuffers 1 and 3, and 100% activity in buffers 2 and 4. NEBuffers 1-4 must be supplemented with 0.1% Triton X-100 or 100 µg/ml BSA to achieve these values.It can also be used in Cutsmart Buffer. To see its % functional activity in Cutsmart, and that of other DNA modifying enzymes in the cloning workflow, refer to the Activity of DNA Modifying Enzymes in CutSmart® Buffer chart.