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Team:TU Darmstadt/Notebook/Labjournal/K1497021
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Latest revision as of 15:17, 17 October 2014
K1497021 - Naringenin Sensor (FdeR) with mKate as Repoter
Another reporter was mKate, which we got out the registry as well (K1055000). We cut the K1055000-plasmid with XbaI + PstI and cut the Naringenin sensor plasmid with SpeI + PstI. In addition, we dephosphorylized the digestion of the naringenin sensor. Subsequently we ligated the insert into the backbone and behind the Naringenin Sensor using T4 ligase, producing a mixed site between the sensor and the reporter. We transformed the ligation product into Top10 E. coli via heatshock and spread them out on CMP-LB agar plates. We incubated the plates overnight at 37°C and analyzed obtained colonies by colony-PCR and agarose gel electrophores. Positive colonies were inoculated in 5 ml LB with CMP and plasmids were isolated after incubating for 16 h at 37°C. For further verification we send the samples to Eurofins for sequencing.
