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Sensing

Results of the gradient PCR of the Sensing constructs from the previous day
The Gradient PCR of the Backbones for the sensning 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 succesful and we used the liearized 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.