Team:RHIT/Protocols/DNADigest

• 
• 
  HOME
• 
  TEAM
  • Team Members
  • Attributions
  • Official Team Profile
• 
  PROJECT
  • Wetlab
  • Human Practices
  • Sustainability
• 
  NOTEBOOK
  • Protocols
  • Journal
  • Parts
  • References
• 
  SAFETY
• 

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.