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iGEM 2010
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Materials & Methods Biosafety Journal Club
bacterial lawn exposed to uv |
Prepare cell lawns for UV exposure. Escherichia coli transformed with either pTSMa or pLPTa and a composite biobrick (one of K415006, K415013, K415022 or K415023) were grown 12-14 hours in 4mL aliquots of LB at 37°C with Kanamycin and Ampicillin, and IPTG to a final concentration 2.65 uM. An OD measurement was taken to confirm the cells to be in the log phase of growth. The remainder of the protocol was performed in the dark. The IPTG was washed out in two cycles of pelleting, decanting, and resuspending the pellet in the original volume of fresh LB. The cells were then concentrated 10x. This sample was added 1:40 to 4mL melted M9 minimal top agar () and kept at 50°C while Kanamycin and Ampicillin were added. If the composite biobrick was not K415022 or K415023, the mixture received M Acyl Homoserine Lactones (AHL) to a concentration of 0.3uM. The mixture was vortexed, quickly poured into small petri dishes, and allowed to cool for 5 minutes. The plates then grew at 30°C for 4 hours. Expose cell lawns to UV. |
