Regulatory

Part:BBa_J100534:Experience

Designed by: Cathleen Krabak, Lindsey Voelker, Kate Kilpatrick, Eboni Thomas   Group: Campbell M Lab   (2020-02-09)


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Applications of BBa_J100534

RFP produced per cell: (luminescence is displayed on the Y-axis)

Graph2020.png

Average RFP produced per cell (luminescence is displayed on the Y-axis):

Simplegrapherror2020.png

PCR gel electrophoresis:

Gelelectro2020.png

User Reviews

UNIQ72065dc13f7013bc-partinfo-00000000-QINU UNIQ72065dc13f7013bc-partinfo-00000001-QINU

Data was derived by dividing fluorescence of each sample by optical density once readings had been collected from the spectrophotometer. The collected data seems to show a general trend of RFP production being hindered by incubating each experimental group in 42 degrees Celsius for one hour. The control experimental groups produced slightly more RFP. This suggests that the heat may have denatured the promoter and reduced its functioning capacity. The original X colonies were derived from red cells produced after we prepared our bacterial cells. Three red cells were produced in the experimental colony, which we used for the X1, X2, and X3 samples. The X1 colony was grown from a cell that was very faintly red; this could account for the fact that little RFP was produced in the presence and absence of heat. Therefore, we decided to exclude the X1 data from the X control and X heat averages on the second graph pictured above. Though the data seems to suggest that the control X group produced more RFP than the heat X group, the error bars on the second graph (which shows the averages) overlap, suggesting that there is no statistically significant difference between the data. Therefore, we are forced to accept the null hypothesis (heat has no effect on RFP production). To improve upon this experiment, we could increase the temperature to see if more significant data is produced, or we could throw out the hypothesis entirely and formulate a new one. The gel electrophoresis revealed relatively successful PCR results. In the left column molecular weights were used for comparison. The next three column include X1, X2, and X3 samples. The right most column contains the negative control sample. The X samples traveled further than the negative control sample, which makes sense because the oligo that we inserted and cloned were smaller (only 60 nucleotides plus sticky ends maximum) than the promoter sequence in the negative control sample that produced GFP. The X1 sample traveled slightly farther than the other two, which aligns with the fact that there may have been some error in the production of that colony.