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Restriction Digestion

  1. Add ddH2O to a tube (usually 20 μL total volume).
  2. Add BSA (provided as 100x concentrate) if necessary.
  3. Add the appropriate restriction digest buffer (provided as 10x concentrate).
  4. Add DNA (plasmid / PCR product).
  5. Add restriction enzyme(s) to the tube. Mix well by pipetting. The volume of restriction enzymes added should be less than 5% of the total volume.
  6. Incubate at the appropriate temperature (usually 37°C) for the required time.

The New England Biolabs website provides useful information on restriction digestion.