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  • Protocol for T7 DNA Ligase (M0318)

    1. Combine 50 ng of vector with a 3-fold molar excess of insert. Adjust volume to 10 µl with dH2O.
    2. Add 10 µl of 2x T7 DNA Ligase Reaction Buffer and mix.
    3. Add 1 µl of T7 DNA Ligase and mix thoroughly.
    4. Centrifuge briefly and incubate at room temperature (25°C) for 15-30 minutes.
    5. Chill on ice, then transform or store at -20°C.
    6. Do not heat inactivate. Heat inactivation dramatically reduces transformation efficiency.