Team:Groningen/Promoters
From 2009.igem.org
This page shows the measurements we did of our constitutive promoters and derives approximate RPUs relative to the weakest promoter we use. For briefness the lines in the graphs use the original numbering, which can be summarized as follows:
Plasmid/Part →  pSB1AC3  pSB3K3  BBa_J23109  BBa_J23106  BBa_J23100  control  
Medium →  LB  TY  LB  TY  LB  TY  LB  TY  LB  TY  LB  TY  
Dilution (of culture) ↓  1  7  2  8  3  9  4  10  5  11  6  12  

100x  A  
B  
200x  C  
D  
400x  E  
F 
Background
The following graphs show the optical density and fluorescence measurements for the controls. As can be seen 4 of the LB medium controls were probably contaminated, so these are ignored in the rest of the computations.
LB controls  TY controls 

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Based on the noncontaminated control measurements we find the following average background values:
 LB
 OD600: 0.094262500378219 (A, F)
 Fluorescence: 11.4090909090909 (A)
 Fluorescence: 20.6136363636364 (B,C,D,E,F)
 TY
 OD600: 0.09115113622763 (AF)
 Fluorescence: 9.06818181818182 (A)
 Fluorescence: 20.05 (B,C,D,E,F)
Using the above background values we get the graphs below.
BBa_J23109  BBa_J23106  BBa_J23100  

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Relative promoter units
Using the computed absorbance and fluorescence values we can determine RPUs (Kelly2009) using the formula:
((d/dt)F_{θ}/ABS_{θ})/((d/dt)F_{J23100}/ABS_{J23100})
Note that we did not test with the official reference promoter and with RFP instead of GFP, as this happened to be easily avaible. In other words, these results should NOT be compared with other computed RPU values, but only be taken as a rough indication of the relative performance of these three promoters.
Reference = BBa_J23100  

BBa_J23109  BBa_J23106  
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Based on the data from t>400min (>24000s) the minimum, maximum and average RPU values were computed (see the "RPU total" sheet in the underlying spreadsheet for the raw values, before rounding):
BBa_J23109  BBa_J23106  

Min RPU  0.011  0.28 
Max RPU  0.033  0.84 
Average RPU  0.015  0.49 
Although the error in the RPU computations is likely to be far from Gaussian, based on the graphs the average provides a reasonable estimate.