For blunting reactions requiring removal of overhangs: T4 DNA Polymerase can be used in NEBuffers r1.1, r2.1, and rCutSmart Buffer as well as NEBuffers 1, 2, and 4 and T4 DNA Ligase Reaction Buffer. NEBuffers r3.1 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 r2.1. BSA Recombinant Albumin (rAlbumin) supplementation is recommended when using a buffer that does not already contain BSA.
To see its % functional activity in rCutsmart, and that of other DNA Modifying enzymes in the cloning workflow, refer to the Activity of DNA Modifying Enzymes in rCutSmart® Buffer chart.