Paris/July 3

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See [[Paris/PROTOCOLS#Preparation of the P1 stock on the w121 strain.|protocols]].
See [[Paris/PROTOCOLS#Preparation of the P1 stock on the w121 strain.|protocols]].
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: ==
+
== Kinetic 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 :
Line 8: Line 8:
=== 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 ===
We measure the kinetic of the w121 in an array <br>
We measure the kinetic of the w121 in an array <br>
-
200µl LB: +0µl DAP, +2µl DAP, +4µl DAP, +8µl DAP, +12µl DAP
+
{{Paris_KineticArray| Title = Growthing of w121 strain as a function of DAP concentration (1µL w121 strain incubated ON)
 +
|A1=|A2=|A3=|A4=|A5=|A6=|A7=|A8=|A9=|A10=
 +
|B1=
 +
|B2=200µL LB + 0µL DAP
 +
|B3=200µL LB + 2µL DAP
 +
|B4=200µL LB + 4µL DAP
 +
|B5=200µL LB + 8µL DAP
 +
|B6=200µL LB + 12µL DAP
 +
|B7=|B8=|B9=|B10=
 +
|C1=|C2=|C3=|C4=|C5=|C6=|C7=|C8=|C9=|C10=
 +
|D1=|D2=|D3=|D4=|D5=|D6=|D7=|D8=|D9=|D10=
 +
|E1=|E2=|E3=|E4=|E5=|E6=|E7=|E8=|E9=|E10=
 +
|F1=|F2=|F3=|F4=|F5=|F6=|F7=|F8=|F9=|F10=
 +
|G1=|G2=|G3=|G4=|G5=|G6=|G7=|G8=|G9=|G10=}}
 +
[DAP] = 5mM
=== The excretion of Dap by MG1655 DapA+ strain by an indirect way : ===
=== 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>
-
200µl S0: +0µl DAP, +2µl DAP, +4µl DAP, +8µl DAP, +12µl DAP
+
{{Paris_KineticArray| Title = Growthing of w121 strain 'supplemented' with S0 (1µL w121 strain incubated ON)
 +
|A1=|A2=|A3=|A4=|A5=|A6=|A7=|A8=|A9=|A10=
 +
|B1=
 +
|B2=200µL S0 + 0µL DAP
 +
|B3=200µL S0 + 2µL DAP
 +
|B4=200µL S0 + 4µL DAP
 +
|B5=200µL S0 + 8µL DAP
 +
|B6=200µL S0 + 12µL DAP
 +
|B7=|B8=|B9=|B10=
 +
|C1=|C2=|C3=|C4=|C5=|C6=|C7=|C8=|C9=|C10=
 +
|D1=|D2=|D3=|D4=|D5=|D6=|D7=|D8=|D9=|D10=
 +
|E1=|E2=|E3=|E4=|E5=|E6=|E7=|E8=|E9=|E10=
 +
|F1=|F2=|F3=|F4=|F5=|F6=|F7=|F8=|F9=|F10=
 +
|G1=|G2=|G3=|G4=|G5=|G6=|G7=|G8=|G9=|G10=}}
-
[DAP] = 5mM (''yes but what does it refer to ? [[User:Nicolas C.|Nicolas C.]])''
+
[DAP] = 5mM
[[Paris|<<home]]
[[Paris|<<home]]

Revision as of 21:00, 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.

Kinetic 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

Kinetic Array :Growthing of w121 strain as a function of DAP concentration (1µL w121 strain incubated ON)
1 2 3 4 5 6 7 8 9 10 11 12
A {{{A11}}} {{{A12}}}
B 200µL LB + 0µL DAP 200µL LB + 2µL DAP 200µL LB + 4µL DAP 200µL LB + 8µL DAP 200µL LB + 12µL DAP {{{B11}}} {{{B12}}}
C {{{C11}}} {{{C12}}}
D {{{D11}}} {{{D12}}}
E {{{E11}}} {{{E12}}}
F {{{F11}}} {{{F12}}}
G {{{G11}}} {{{G12}}}
H {{{H1}}} {{{H2}}} {{{H3}}} {{{H4}}} {{{H5}}} {{{H6}}} {{{H7}}} {{{H8}}} {{{H9}}} {{{H10}}} {{{H11}}} {{{H12}}}

[DAP] = 5mM

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

Kinetic Array :Growthing of w121 strain 'supplemented' with S0 (1µL w121 strain incubated ON)
1 2 3 4 5 6 7 8 9 10 11 12
A {{{A11}}} {{{A12}}}
B 200µL S0 + 0µL DAP 200µL S0 + 2µL DAP 200µL S0 + 4µL DAP 200µL S0 + 8µL DAP 200µL S0 + 12µL DAP {{{B11}}} {{{B12}}}
C {{{C11}}} {{{C12}}}
D {{{D11}}} {{{D12}}}
E {{{E11}}} {{{E12}}}
F {{{F11}}} {{{F12}}}
G {{{G11}}} {{{G12}}}
H {{{H1}}} {{{H2}}} {{{H3}}} {{{H4}}} {{{H5}}} {{{H6}}} {{{H7}}} {{{H8}}} {{{H9}}} {{{H10}}} {{{H11}}} {{{H12}}}

[DAP] = 5mM

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