Team:UPO-Sevilla/Notebook/09 29


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September, 29th

Assay Team

We have analyzed plates spread in the previous chemotaxis assay. There were no colonies coming from non succinate cubicles in those plates. That could mean that P. Putida KT2442 needs succinate as an energy source to show chemotaxis behaviour. We wanted to observe if the results were reproducible and in this case they were not.Needles are supposed to show a similar number of colonies when having the same content and being in the same cubicle. Yet only in the 10mM Asp + Succ cubicle that would happen. We had to continue working on the design of the chemotaxis assays to achieve reproducible results. Finally we could observe that there was a tendency to increase the number of bacteria in needles when using aspartate (in cubicles with succinate). Nonetheless, owing to the non reproducible results, we could not admit that as a success.

We have checked our devices and vectors to demonstrate if these are correct and we have enough concentration for our final circuits. Different plasmid preparations were quantified by Nanodrop. All of them had good concentration, but UPO13+3. Digestion of 1+2; 12+2;13+3;16+3;7+3; 12+19; 1+2+16+3; 1A3; 1C3; 1AC3; 1AT3; 4K5; 3C5; 3T5; 4C5; 1K3 (2h, 37ºC). These digestions were run in a 0.8% or 1.5% agarose gel. The following samples did not correspond to expected size: 7+3; 13+3; 1+2; 12+2.

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