IGEM:UNAM/2009/Notebook/Modeling logbook Claudia/2010/08/20

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Working on LovTAP promoters: Restrictions of Medium and Weak Promoters
Once the cells were correctly transformed with the six plasmids harboring the  three weak and three medium promoters, I have started the plasmid extraction procedure.



Plasmid extraction Protocol
I am using the High Pure Plasmid Isolation kit from Roche

Restriction enzyme assay:Preparation for ligation
After finishing the isolation of the six plasmids, I have started the restriction enzyme assay in order to remove the RBS site, the RFP gene and the doble terminator from each one to replace them with LovTAP gene.


 * Restriction enzymes Mixture:

Restriction enzymes: SpeI and PstI.

The restriction reactions were incubated at 37°C overnight.

Note: Inactivate the enzymes at 80°C during 10 min.


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