Part 1 – The PCB
Fig. 1 - 3D Model of the PCB
Fig. 2 - XOR curve on the scope.
Fig. 3 - XOR curve on the scope.
Fig. 4 - PCB Pinheader Marked.
Fig. 5 - Matlab Plot of the derived phase shift and time shift in 2D and 3D illustration.
In conclusion, it can be said that even though there have been a number of systematic measurement deviations, we have developed a functional evaluation board that allows us to determine a phase shift from the analog output voltage value.
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Part 2 – Engineering of FbFPs
After a long research for a FbFP candidate, we found the protein SB2 from Pseudomonas putida, which shows a measurable fluorescent activity. So, we designed a testing system to measure the fluorescent activity of the SB2 protein (BBa_3410007) in contrast to the FbFP protein available in the parts registry of iGEM (BBa_K3013002). For this purpose, we chose the vector pJoe as our backbone and cloned the testing construct in silico with SnapGene (figure 6).
Figure 6: Plasmid map of the vector pJoe with the insert SB2
Figure 7: Gel image for control of the PCR of the SB2 protein, the registry part and the vector pJoe.
Figure 8: Colony PCR from SB2, the registry part and protein KOFP-7 in three different gels
Figure 9: Results of fluorescence measurement after 10 to 16 hours after induction with IPTG in the Tecan reader