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.

Pae 8 jul 2011.JPG This gel comes from here

The transformation was did according to this protocol