Lidstrom:Digestion with Restriction Enzymes
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- Try it quick and imprecise like iGEM 2011 UW recommends:
- 10 uL DNA (totaling 500+ ng is good)
- 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.
- 1 - 2 hours at 37oC or room temperature overnight
- Inactivate by ..