Regulatory

Part:BBa_J100180:Experience

Designed by: Anna Buser, Jerry Chang, Natalie Philips, Ali Kamal   Group: Campbell M Lab   (2014-09-11)
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In our experiment, we tested our P2(2) promoter against a positive control (lacI promoter), and a negative control (backwards promoter). Additionally, we tested each promoter at three pH levels (6.0, 7.0, 8.0) to see the effect of pH on the promoter.

Cells with experimental P2(2) were grown overnight in buffer of pH 6.0, 7.0, and 8.0. The same was done for cells with the positive control (lacI promoter) and negative control (backwards promoter). Samples of each promoter from each pH level were loaded into a spectrophotometer to measure red fluorescence divided by cell density. Buffer was also measured in the spectrophotometer to act as a baseline reading, which we subtracted from each trial. Averages of fluorescence/density are shown on the graph; error bars depict standard deviation.

The first graph shows successful transformation with the positive control (lacI) at all three pH levels with a significant drop in transformation in both the negative control and the experimental P2(2). The second graph compares our experimental P2(2) promoter against the negative control, and shows no significant difference between the two.

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