Lidstrom:Digestion with Restriction Enzymes

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Recipe

• Try it quick and imprecise like iGEM 2011 UW recommends:
  • 10+ uL DNA (
    • Want at least 500 ng. Can elute a miniprep in 38 uL of water and use that whole thing. Don't use miniprep DNA eluted in buffer; only use DNA eluted in water.
  • 5uL buffer (check http://www.neb.com/nebecomm/EnzymeFinderSearchByName.asp for a list of enzymes and associated buffers)
  • 0.5 uL BSA (if required. See above)
  • 1uL enzyme 1
  • water to 50 uL (32.5 uL, add first)
  • If two enzymes are required, check http://www.neb.com/nebecomm/DoubleDigestCalculator.asp for buffer, temperature, BSA specifications.

• If using BioBrick or BglBrick enzymes, chose your buffer based on Janet's notes.

Temperature/Time

• 1 - 2 hours at 37oC or room temperature overnight
• Inactivate by ..