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  • FAQ: Can T4 DNA Polymerase be used in other NEBuffers and CutSmart?

    For fill-in reactions only: T4 DNA Polymerase can be used in NEBuffers 1.1, 2.1, 3.1 and CutSmart® Buffer as well as NEBuffers 1-4 and T4 DNA Ligase Reaction Buffer.

    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.