Restriction Digest Add the following reagents to a tube Reagent50 ul reaction10 ul reaction Cutsmart buffer * 5 ul 1 ul Enzyme 1 10 units 1 unit Enzyme 2 10 units 1 unit DNA 500 ng 100 ng Water to 50 ulto 10 ul * Note: if not using EcoRI-HF, PstI-HF, SpeI-HF, and XbaI, a different buffer by be needed. Consult the NEB performance chartfor more information Incubate for 1 hour at 37C Heat inactivate for 20 minutes at 80C This step is only necessary if the reaction will not be used immediately
