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Restriction Enzymes M.3: RESTRICTION ENZYME DIGESTION OF DNA                                         Go Home


1. Add the following to a microfuge tube:
2 ul of appropriate 10X restriction enzyme buffer
0.1 to 5 ug DNA
sterile water to a final volume of 19 ul
2. Add 1 to 2 ul (3 to 20 units) enzyme and mix gently. Spin for a few seconds
in microfuge.
3. Incubate at the appropriate temperature (usually 37oC) for 1 to 2 hours.
4. Run a small aliquot on a gel to check for digestion.
5. If the DNA is to be used for another manipulation, heat inactivate the enzyme (if it is heat labile) at 70oC for 15 min or phenol/chloroform extract and

ethanol precipitate.