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  • Quick Ligation Protocol (M2200)

    Buffers

    Quick Ligation Reaction Buffer

    Protocol

    1. Combine 50 ng of vector with a 3-fold molar excess of insert. Use NEBioCalculator to calculate molar ratios. Adjust volume to 10 μl with dH2O.
    2. Add 10 μl of 2X Quick Ligation Buffer and mix.
    3. Add 1μl of Quick T4 DNA Ligase and mix thoroughly.
    4. Centrifuge briefly and incubate at room temperature (25°C) for 5 minutes.
    5. Chill on ice, then transform or store at -20°C.
    6. Do not heat inactivate. Heat inactivation dramatically reduces transformation efficiency.