For blunting reactions requiring removal of overhangs: T4 DNA Polymerase can be used in NEBuffers 1.1, 2.1, and CutSmart Buffer as well as NEBuffers 1, 2, and 4 and T4 DNA Ligase Reaction Buffer. NEBuffers 3.1 and 3 are not recommended when overhang removal is required.
Supplementation of dNTPs is required for all blunting reactions. Optimal activity is observed in NEBuffer 2.1. BSA supplementation is recommended when using a buffer that does not already contain BSA.
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.