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
BfaI NotI SphI-HF
BspCNI PacI StyI
BspEI PvuI StyD4I
BtgZI RsrII Tsp45I

There is also a protocol using two RE-Mixes.