Optimal activity is observed in Isothermal Amplification Buffer and near optimal performance is observed in ThermoPol™ reaction buffer. The unit is assayed in a non-optimal buffer taken from the literature. The enzyme exhibits 25% activity in NEBuffer 1 and 100% activity in buffers 2, 3, 4, and EcoRI. NEBuffers 1-4 should be supplemented with 0.1% Triton X-100, 0.1% Tween-20 or 100 µg/ml BSA to achieve best results.