Paris Notebook
From 2007.igem.org
Monday, July 2
First day in the lab ! - Overview of the project - Planning of the lab work - Primer design
We got the W121 strain from a lab in Pasteur Institute. This strain is [DapA-; Erythromycin R], but also has a couple of other mutations we are not interested in. Thus we will need to do a transduction of the deletion to the strain we will use: MG1655
We launched ON culture of w121 to prepare a stock of P1 phages.
In order to model the dynamics of the synthetic organism, we need to measure several parameters. Two of which are: - The growth of the DapA- strain relative to the DAP concentration of the medium - The excretion of DAP by the DapA+ strains
The first one is easy, we just need to measure growth kinetics of our DapA- strain in function of the concentration of the DAP we add in the medium. The second one is more tricky. Direct dosage of DAP is quite complicated and expensive, thus we will try a kind of bio-measurement. We will grow the DapA+ strain ON, then we will centrifugate to recover the culture medium. We will filter this culture medium to sterilize it and we will grow a DapA- strain in it. The growth curve should enable us evaluating the medium concentration in DAP.
Tuesday, July 3
- Preparation of the P1 stock on the w121 strain. See protocols.
- Kinetics measurements:
200ul LB: +0ul DAP, +2ul DAP, +4ul DAP, +8ul DAP, +12ul DAP
200ul S0: +0ul DAP, +2ul DAP, +4ul DAP, +8ul DAP, +12ul DAP
S0= Centrifuged and filtered medium of MG1655 ON DAP is 5mM
