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  • Double Digest Protocol using One RE-Mix and One Standard Restriction Enzyme


    1. Dilute up to 1 μg DNA to 17 μl with dH2
    2. Add 2 μl of the 10X RE-Mix and 1 μl of the standard enzyme
    3. Incubate at 37°C, for 15 minutes Time-Saver enzymes, or 1 hour for standard enzymes
    4. Analyze by agarose gel electrophoresis
    Note: Use only with standard restriction enzymes with 37°C incubation temperature.

    Some Standard Restriction Enzymes are not compatible with RE-Mix

    Standard Restriction Enzymes Not Compatible with RE-Mix Master Mixes
    (Sequential Digestions Recommended)
    AleI DpnII SalI
    AlwI MluI SexAI
    BanI MwoI SfaNI
    BciVI NmeAIII SgrAI
    BfaI NotI SphI-HF
    BspCNI PacI StyI
    BspEI PvuI StyD4I
    BtgZI RsrII Tsp45I

    There is also a protocol using two RE-Mixes.