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Sensing

DpnI digest of the iGEM promoter and the GelE Backbone
I digested the newly amplified and purified PCR products. The concentrations were not too good but I went ahead with the Gibson Assembly.

Miniprep of the Cad-transformants&PCR thereof
I did the MIniprep of the innoculated cad Transformants and then I did a 20ul reaction PCR in order to check for the presence of the Cad promoter insert. The PCR showed that it was indeed present.

Gibson Assembly of the constitutive promoter+GFP (sensing) and the GelE+GFP construct (effector)
I assembled the constitutive promoter in the plasmid in front of GFP so that we could use it as a positive control for the sensing module. I also inserted the GelE gelatinase gene into the plasmid containing the arabinose sensitve promoter.

Colony PCR of the MMP9 construct
In order to check for the presence of the MMP9 insert I did a colony PCR with the colonies that were transformed with this plasmid. But the colony PCR did not work so I decided to innoculate the colonies and do the PCR on the MiniPrep the next day.

Transformation of cells with the constitutive promoter+GFP and the GelE gelatinase gene
I transformed competent cells with the two last constructs, the one that would serve as a positive control for the sensing module (constitutive Promoter+GFP) and the other one with the gelatinase gene for GelE.

Note
I also realized that the construct for the effector module would not express GFP because due to the primers there was a frame shift that induced a stop codon just after the linker and before GFP.

Nanoparticles

DLS measurements.