Team:RHIT/Protocols/DNADigest

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DNA Digest

DNA Digestion of the Blue Construct and pRS416 with EcoRI and SpeI


To a 1.5 mL microcentrifuge tube, add the following:

  • 7 µL ddH2O
  • 1 µL 10X cutsmart buffer
  • 1 µL miniprepped DNA
  • 1 µ EcoRI enzyme


Incubate the tube for 1-2 hours at 37°C, and then heat inactivate the enzyme by placing the tube in a water bath at 65°C for 20 min.


After removing the tube from the water bath, repeat the first step, instead adding SpeI as the enzyme. Incubate the tube for 1-2 hours at 37°C. Remove tube from the incubator and place it in a -20°C freezer until the product is to be analyzed by agarose gel electrophoresis.