IGEM:UNAM-Genomics Mexico/2009/Notebook/H2 Pae/2011/07/11

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bjectives 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