Lidstrom:QuikChange Site-Directed Mutagenesis

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Revision as of 09:55, 12 June 2014 by Janet B. Matsen (Talk | contribs)
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QuikChange Basics

  • You can buy a kit from Agilent or use your own ingredients.
  • The marketing term "lightning" refers to an improved Dpn1, that degrades template (unmutated template) more efficiently.

Two (main) QuikChange kits are available

  • QuikChange Lightning.
    • For mutations contained within a single primer
  • QuikChange Lightning Multi
    • For mutations contained in >1 primer
    • Can be used for mutations contained in 1 primer, but this kit is more expensive than the single kit and likely has lower efficiency.
  • Both kits:
    • come with chemically competent XL10-Gold. This is perhaps the most valuable part of the kit.

Tips

  • Though the lightning Dpn1 enzyme claims to be effective in 5 minutes, you might as well incubate it for 30 minutes, just in case it lowers your unmutated background further.
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