IGEM:UNAM-Genomics-Mexico/2009/Notebook/iGEM 2011 UNAM-Genomics Mexico/2011/06/02
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We 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).
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)