-We have made tryptone broth, which will be used to grow E. coli (K-12 D10b strain) overnight before the chemotaxis experiments, for them to develop flagella and therefore they will be capable of chemotaxis. To make tryptone broth look at protocols.
-Cells have been transformed with backbone plasmid pSB1C3 carrying biobrick BBa_K398500, with constitutive promoter J23100, protocol for transformation can be found here.
Chapter X
3rd of August
For experiments involving modified E. coli, we will need positive and negative control. In order to set up a positive control, E. coli expressing GFP is used for observing endogenous chemotaxis system of E. coli. After growth and observation under microscope we have found out that K-12 DH10b does not develop flagella and therefore can not move. Due to this reason we are using another strain, BL21 DE3 which is motile. These bacteria however had no plasmid and to observe them under wide field microscope a flourescent expression is necessary. Due to this we have transformed BL21-DE3 strain with a plasmid containing ampicillin and kanamycin resistance (AK3 backbone).
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