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Tuesday, June 10th

Preparation of -SOC -TFB1 -TFB2 (pH was set to 5.6 instead of 5.8) -Agar plates (containing ampicillin (200 mg/L) or chloramphenicol (34 mg/L)) According to qiagen DNA protocols & application Modifications: -150 ml Glycerol per 1 L (15%) are added to TFB1 and TFB2 (instead of 15 mL) -1.4 g/l CaCl2 dihydrate are used in TFB1 -11.0 g/l CaCl2 dihydrate are used in TFB2 -SOC is sterilized by filtration (not autoclaved)

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Week 6: Project planning, Gibson assemblies planning, First Matlab simulations

Wednesday, July 2nd

• We have a clear organization of the different experiments we would like to perform.

We want to optimize quorum sensing to have non-leaky non-cross-talking constructs. We want to prevent cross-talk between integrases and check their dynamics with different quorum sensing promoters. Then we will test the XOR gate without production of LuxI, to check how it works without the loop, and we will finally test our final construct with the loop, hoping that the delay between GFP production and the possible second switching due to AHL production is long enough to have enough fluorescence.

The red line is our critical timeline. Numbers of days are very optimistic,therefore we multiply the whole timeline by 2. The stars stand for modeling inputs.

Tasks organization with critical timeline in red.

• We have also clear plans for the Gibson assemblies to perform. As an example, here is the plan for regulator plasmids :

Exemple of Gibson assembly plan for regulator constructs

• We have a facebook page !

• We simulated quorum sensing without leakiness on Matlab, with parameters from the literature.