Maheshri:Digestion

 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.