Materials and Methods

The parts for the three constructs used in the InterLab study (J23101, J23106, J23117 and I13504,) were taken from the iGEM distribution plates and transformed into NEB turbo cells. Afterwards, the plasmids were recovered through miniprep from an over night culture. The plasmids were amplified via PCR and purified by gel extraction. The final constructs were then assembled using CPEC cloning (Circular Polymerase Extension Cloning). The correct identity of the resulting constructs was confirmed by analytical digest with PvuII and sequencing. The final plasmids were transformed into DH5α cells. For the inoculation and cultivation we followed the recommended InterLab Protocol. We prepared our devices and the controls by streaking them out on agar plates from glycerol stocks. We incubated the plates 18h overnight at 37°C. For that we used LB Agar from Roth without antibiotic for the empty DH5alpha cells and LB Agar from Roth with Chloramphenicol (c=34 ug/ul) for the remaining strains carrying the constructs and controls. We inoculated our strains carrying the devices and controls in 5 mL LB with Chloramphenicol in a 15 mL Test tube. The empty DH5alpha cells were inoculated in 5 mL LB without antibiotic. For each sample we set up three biological replicas. The samples were incubated at 37°C at 300rpm for 16 h.