Team:TU Darmstadt/Notebook/Labjournal/K1497008

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Revision as of 15:00, 17 October 2014

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K1497008 - hokD (relF)

To generate the hokD-BioBrick we designed primers (relF_Pre_F and relF_Suf_R) that amplify the hokD gene from the genome of E. coli MG1655 via PCR including the BioBrick prefix and suffix. The length of the PCR product was controlled via gel-electrophoresis using a 0.8% agarose gel. The PCR product was cut with EcoRI and PstI and then ligated into pSB1C3, cut with EcoRI and PstI. After transformation into E. coli TOP10 and after 24 hours of incubation the plasmid was extracted via Wizard PCR Clean Up Kit (Promega) and the DNA sequence was verified by Eurofins.

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