Team:TU Darmstadt/Notebook/Labjournal/K1497019
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K1497019 - Naringenin Sensor (FdeR)
We kindly got the Naringenin Sensor from Dr. Siedler. We tested its functionality and because we were not allowed to change anything within the device's sequence, we ordered the sequence as disered. We amplificated the insert via PCR using "PRIMERs" and checked PCR by agarose gel electroporesis. Afterwards we cleaned up the PCR product by DpnI digestion and using the Wizard PCR Clean Up Kit. We cut the clean PCR product with EcoRI + PstI and ligated this part with a previously digestion of mRFP-pSB1C3 with EcoRI +PstI. We transformed the ligation in Top10 via heat shock and spread out on CMP-LB agar plates. After incubating the plates overnight at 37°C, we analyzed white colonies by colony-PCR and agarose gel electrophoresis. Positive colonies were inoculated in 5 ml LB with CMP and plasmids were purified after incubating for 16 h at 37°C. For further verification we send the samples to Eurofins for sequencing.