Part:BBa_J100029:Experience

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Applications of BBa_J100029

In tube 1, we placed the rpoD promoter ligated into the E.coli plasmid without being induced. In tube 2, we placed the rpoD promoter ligated into the E. coli plasmid and induced by heat shocking the promoter overnight at 40 degrees Celcius compared to 37 degree Celcius at which the other 5 tubes were incubated. The negative control was in tube 3, and the plasmid in this tube did not contain a promoter. Instead, it had a transcription terminator which prevented RFP from being produced. Our positive control in tube 4 was a pTet promoter that is constitutive, meaning this particular promoter is always active. In the fifth tube, the plasmid contained the pLac promoter that was induced by a 0.2% concentration of IPTG. And in tube 6, the plasmid contained the pLac promoter that was not induced by the IPTG. Because our experimental promoter in the first tube was not induced, there was a very small amount of fluorescence produced, similar to that produced by the negative control. But the rpoD promoter that was induced by heat shock was able to turn on the gene for RFP and produce a significant amount of fluorescence as a result.

User Reviews

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