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This preparation is necessary for transduction of DapA deletion in MG1655 strain. | This preparation is necessary for transduction of DapA deletion in MG1655 strain. | ||
== Kinetics measurements: == | == Kinetics measurements: == | ||
- | As previously described, we want to measure | + | As previously described, we want to measure : |
* Growth of DapA- strain (w121) relative to the concentration of DAP in the medium : | * Growth of DapA- strain (w121) relative to the concentration of DAP in the medium : | ||
+ | * The excretion of Dap by MG1655 DapA+ strain by an indirect way | ||
+ | === Growth of DapA- strain (w121) relative to the concentration of DAP in the medium === | ||
+ | We measure the kinetic of the w121 in an array <br> | ||
200µl LB: +0µl DAP, +2µl DAP, +4µl DAP, +8µl DAP, +12µl DAP | 200µl LB: +0µl DAP, +2µl DAP, +4µl DAP, +8µl DAP, +12µl DAP | ||
- | + | === The excretion of Dap by MG1655 DapA+ strain by an indirect way : === | |
We measure the growth of DapA- strain in the filtered growth medium of previously incubated MG1655 strain (S0) | We measure the growth of DapA- strain in the filtered growth medium of previously incubated MG1655 strain (S0) | ||
S0 = Centrifuged and filtered medium of MG1655 ON <br> | S0 = Centrifuged and filtered medium of MG1655 ON <br> |
Revision as of 15:33, 10 July 2007
Contents |
Preparation of the P1 stock on the w121 strain.
See protocols. This preparation is necessary for transduction of DapA deletion in MG1655 strain.
Kinetics measurements:
As previously described, we want to measure :
- Growth of DapA- strain (w121) relative to the concentration of DAP in the medium :
- The excretion of Dap by MG1655 DapA+ strain by an indirect way
Growth of DapA- strain (w121) relative to the concentration of DAP in the medium
We measure the kinetic of the w121 in an array
200µl LB: +0µl DAP, +2µl DAP, +4µl DAP, +8µl DAP, +12µl DAP
The excretion of Dap by MG1655 DapA+ strain by an indirect way :
We measure the growth of DapA- strain in the filtered growth medium of previously incubated MG1655 strain (S0)
S0 = Centrifuged and filtered medium of MG1655 ON
200µl S0: +0µl DAP, +2µl DAP, +4µl DAP, +8µl DAP, +12µl DAP
[DAP] = 5mM (yes but what does it refer to ? Nicolas C.)