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Cell Surface DisplayBBa_K523013 Characterization
Characterisation of the iGEM plasmid BBa_K523013 we decided to use.
Sensing-Effector
Making a glycerol stock
-From each culture that was inoculated we made a 50% glycerol stock of 500ul Bacterial culture and 500ul 50% glycerol.
PCR
We Started the Gibson assembly of four of our constructs by making a PCR of each, the Insert as well as the Backbone, making eight different PCR tubes in total (see protocol PCR). About twenty minutes before the PCR reaction was done, the machine malfunctionned. We did run a 0.8% gel, but The results cannot be well interpreted, as the reaction could not go to completion.
We re-ran another PCR reaction with the same constructs, as well as a reactant control. After the PCR the amplified DNA was stored over night at 4°C.
Nanoparticles
Making Nanoparticles, 2nd try
- We washed the nanoparticles with 75% acetone.