Lidstrom:QuikChange Site-Directed Mutagenesis: Difference between revisions

Revision as of 06:55, 12 June 2014

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.

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.

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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 Back to [[Lidstrom:Protocols|Protocols]]
+==QuikChange Basics ==
+* You can buy a kit from Agilent or [[Richard_Lab:Site_Directed_Mutagenesis|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.