IGEM:UNAM-Genomics Mexico/2009/Notebook/H2 Pae/2011/07/11
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To see if our PCR was correctly inserted into pBBRMCS-5, doing a restriction with the same enzymes used to generate sticky end between the plasmid and the PCR, these enzymes are apaI and sacI.
Transformation of pBBRMCS-5
Because apaI doesn´t work with dcm methylase which is present in sp. DH5alfa, I transformated the plasmid into sp. BL21 whic doesn´t have dcm. Tomoorrow I will do the isolation from the colonies resulting from today transformation and the double digestion by apaI and sacI.
I did 3 transformation reactions:
1. DNA taken from the second lane.
2. DNA taken from the fifth lane.
3. No DNA.
This gel comes from here
The transformation was did according to this protocol