Part:BBa_K270003:Experience

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Applications of BBa_K270003

UTDallas 2010 Experience

• The data was taken using E. coli DH5α that was co-transformed with Pu+GFP and Pr+XylR. This was then grown overnight in 2mL of LB broth that had 50ug/mL carbenicillin and kanamycin. This was used in a 1:50 dilution. Each sample had 40uL from the broth grown overnight and 2mL of LB carb. and kan. These were allowed to grow until a predetermined OD. Then the aromatics were added and the cells were allowed to grow for another 2 hours, before the images were taken.
• All images were taken with Olympus IX81 automated inverted microscope specially equipped for live cell imaging. All microscope images were taken from live Dh5a E.coli cells. The filter sets we used are:470/40x (excitation) and 525/50m (emission) for GFP. Data collection and processing was performed by the SlideBook software.
• The leftmost image is the negative control. The other six images are the other concentrations (10uM and 500uM) in the specific aromatic: xylene, benzene, and toluene. The graph is of the average of all the maximum intensities of the cells in the picture vs the concentration of the aromatic added to the sample. We also took data from 10mM and 100mM concentrations of aromatics, but most of the cells died at these higher concentrations.

User Reviews

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