IGEM:UNAM-Genomics-Mexico/2009/Notebook/iGEM 2011 UNAM-Genomics Mexico/2011/06/02: Difference between revisions
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* T4 ligase -> 1 μL | * T4 ligase -> 1 μL | ||
Leave the reaction for two hours at 25ºC | For two total reactions of 10 μL. | ||
Leave the reaction for two hours at 25ºC | |||
====Transformation into E. coli DH5α==== | ====Transformation into E. coli DH5α==== | ||
Revision as of 07:52, 10 June 2011
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Digestion ControlWe ligated the product of the digestion to make sure the plasmid PBBRMCS-5 was indeed digested with both enzymes (XBA I and Hind III). Then we transformed the plasmid with E. coli DH5α. We expect that the transformation will not yield many E. coli colonies (there is not a perfect digestion). Ligation
Ligation Control
For two total reactions of 10 μL. Leave the reaction for two hours at 25ºC Transformation into E. coli DH5α
* Tube 1 -> Transformation Control (no Plasmid DNA added)
* Tube 2 -> 4 μL of ligated plasmid DNA added
* Tube 3 -> 4 μL of the ligation control added (no plasmid DNA added)
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