Team:EPF Lausanne/Calendar/4 September 2013


Taxi.Coli: Smart Drug Delivery iGEM EPFL


Cell Surface Display
Plating of SD and SI from glycerol stock.
Colony PCR of SA gibson transformed cells.


Results of the gradient PCR of the Sensing constructs from the previous day
-The Gradient PCR of the Backbones for the sensing constructs did not work. So we wanted to digest the backbone with an enzyme that cuts in the region of the AraC promoter, as this part would be removed during later PCR for the sensing constructs. This would yield a linear DNA which would be easier to amplify.

Restriction digest
-We digested the plasmid with the BamHI enzyme which cuts it in the AraC promoter. The digestion was successful and we used the linearized plasmid to do another PCR with the primers for the hya backbone.

PCR of the three sensing Backbones with a different program
-We tried a new program which runs the first ten cycles 10°C bellow the normal annealing temperature and then the 20 remaining ones at the normal temperature. Finally this PCR worked and the Backbones were amplified.

-We decided to only do the effector constructs that contained GFP because we did not have time to do them all. Also, it would be easier to check if the promoter is really still induced by arabinose or not.


Sensitivity Assay
-DLS sensitivity assay (i.e. measuring diluted nanoparticles to see how far we could go in the dilutions)
- start of a new ELISA-like assay