From 2007.igem.org

First day in the lab !

Contents 1 Overview of the project 2 Planning of the lab work 3 Primer design 4 Preliminary work on w121 strain 4.1 Transduction of DapA- deletion from w121 to MG1655 4.2 Measuring kinetic parameters of our strains

Overview of the project

Planning of the lab work

Primer design

Preliminary work on w121 strain

Transduction of DapA- deletion from w121 to MG1655

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 (= OverNight) culture of w121 to prepare a stock of P1 phages.

Measuring kinetic parameters of our strains

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+] strain.

The first one is easy, we just need to measure growth kinetics of our [DapA-] strain contingent on 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.

TEST TEST2