IGEM:Virginia/2012/Notebook/Genetically engineered bacteriophage for diagnosis of whooping cough/2012/08/26

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Week of 8/26/12

• Transformed BBa_K331022 (which will be used for the pSB1C3 backbone), BBa_K091110 (which will be used for expressing genes under the control of the lacI promoter), and the SynCG plasmid into DH5α and grew them overnight on plates. The BBa_K331022 and SynCG plasmids grew on the plates. These were then grown up in a liquid culture and plasmid prepped, resulting in 43.7 ng/µL
• Digested bbp9 (E+S), bbp11 (E+S), promoter+RBS (E+S), BBa_K133022 (E+S), native hCG (E+S), SynCG plasmid (XbaI), and T7 genome (BclI). Induced culture of DH5α of full hCG in vector with IPTG. The bbp9, bbp11, K331022, and WT hCG produced suitable amounts after gel extraction. However, the promoter+RBS needs to be redone and purified with ethanol precipitation along with the SynCG and the T7 genome.