Team:SDU-Denmark/Tour54
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Interlab study
Figure 1:
We have participated in the iGEM interlabstudy.
The focus of this study were to characterize three different promoters, using a gene encoding GFP.
GFP is the reporter protein of choice, as it has a low folding time, and can be quantified through fluorescence, instead of RNA.
Our biobricks were assembled through standard biobrick assembly.
The three constructs were made through standard biobrick assembly, inserting GFP into plasmids containing different promoters. The final construct were transformed into E. coli K12 TOP10.
Figure 2:
Once the final constructs were made, each construct were veryfied through colony PCR, and sequencing.
For the measurements conducted in this project we used FACS.
FACS functions through singling out droplets containing cells, exciting the proteins by shooting light on the absorption frequency onto the cells, then by measuring the emission intensity, the fluorescence of each cell is measured.
When measuring we used our own construct rfirefhyiw as a positive control, and TOP10 as negative control.
Each construct were in the experiment measured in both biological and technical triplicates.
Although veryfied through colony PCR, one construct seemed faulty, due to irregular measurments, and inconclusive sequencing.