Kick off event 19.06.13

Brainstorming 19.06.13 - 3.07.13

Coming up with different ideas for our iGEM project, considering the advantages and disadvantages and the final impact. In the end, we decided on something innovative and fun!

Start Work 4.07.13

Week 1:

PLANNING AND LAB WORK.
Planning of responsibilities and establishment of roles of all team members; starting the wiki; gathering more information on the project by doing research in literature; designing a logo for our product, coming up with a catchy slogan, and deciding on a name for the final project; designing initial experiments and learning many necessary lab techniques (some of the first lab tasks included making buffers, media, chemical competent cells, choosing biobricks for our cells, and doing transformations of the first chosen bricks.

Week 2:

CONTINUATION OF LAB WORK AND BEGINNING OF MODELING.
Transforming all the biobricks and cloning to build our chosen pathways; after research in literature, deciding on 4 hydrolases : NagZ, PhoA, GusA and Aes as well as respective substrates to color them.
We encountered some difficulties during transformation of triple knockout cells with the hydrolases, so this step will have to be repeated in the following weeks with a different strain of bacteria.

Week 3-4 :

MORE LAB WORK AND MODELING.
Transforming and cloning; designing primers for the mutation of LuxR promoter to alter sensitvity; starting AHL diffusion experiments for characterization of the AHL diffusion.
The receiver cells react to AHL in liquid culture and on Agar plates (proof of principle for the receiver cells)

Week 5-6 :

LAB WORK, MODELING, UPDATING WIKI.
Working on AHL diffusion, particularly its efficacy; transforming a new strain of E.coli with the T7 polymerase to test the substrates; working on transforming the triple knockout cells; retransforming the LuxI (K805016) construct because the first one (C0061) din't have an RBS.

Week 7 Half-time:

LAB WORK, MANY UPDATES ON THE WIKI, MODELLING, WORKING ON HUMAN PRACTICES.
Great pictures for the AHL diffusion on single layer agar with GFP receiver cells; Pcons-LuxI (strong promoter) was transformed. The supernatant of the Pcons-LuxI overnight culture induces GFP in the receiver cell (proof of principle for AHL quorum sensing of sender and receiver cells);linear titration of the AHL concentration.PCR of the hydrolases for the biobricks.Do random mutagenisis of the Plux promoter for different affinities.The pLuxR mutation worked and we saw different fluorescent responses to an AHL concentration of [5uM].First Model created about the AHL diffusion on Agar.

Week 8:

LAB WORK , WIKI UPDTAES, HUMAN PRACTISES, MODELING.
Mutagenized promoter selection by comparing the sensivities to the sensivity of the Wild-Type pLuxR promoter. Sender cell receiver cell experiment prooves that quorum sensing, as well as our sender and receiver cells work properly