Paris/July 24

From 2007.igem.org

(Difference between revisions)
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<bbpart>BBa_E0241</bbpart> in <bbpart>pSB1A2</bbpart> - Clone 1 - MP7.1'<br>
<bbpart>BBa_E0241</bbpart> in <bbpart>pSB1A2</bbpart> - Clone 1 - MP7.1'<br>
<bbpart>BBa_J61047</bbpart> in <bbpart>pSB1A2</bbpart> - Clone 1&2 - MP9.1' & MP9.2'<br>
<bbpart>BBa_J61047</bbpart> in <bbpart>pSB1A2</bbpart> - Clone 1&2 - MP9.1' & MP9.2'<br>
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== Growth kinetics of w121 strain ==
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We make an array to test growth of w121 on different growth media (LB, S0.2, S0.4, S0.6, S0.8), supplemented with different amount of DAP.
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Two questions are addressed by the following assay:
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-What is the growth behaviour of w121 (dapA- strain) at different concentrations of DAP?
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-How does w121 strain grow on filtrates of MG1655 growth medium; that is, does MG1655 secrete DAP during growth?
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MG1655 was grown on LB medium and the growth medium was filtered free of bacteria at different DO (Optical Densities) during exponential growth phase:
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S0.2 (at DO=0.2) 
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S0.4 (at DO=0.4)
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S0.6                       
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S0.8                       
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W121 was grown on different media & Growth kinetics were measured:
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LB                          line B in the array
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S0.2 (at DO=0.2)            line C in the array
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S0.4                        line D in the array
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S0.6                        line E & F in the array
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S0.8                        line G in the array
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In the different columns, DAP was added to the indicated final concentrations (without taking into account DAP produced by MG1655 regarding the recycled growth media)
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{{Paris_KineticArray| Title = w121 kinetic as a function of DAP and supplemented medium (S0.x)
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|A1=H<sub>2</sub>0|A2=H<sub>2</sub>0|A3=H<sub>2</sub>0|A4=H<sub>2</sub>0
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|A5=H<sub>2</sub>0|A6=H<sub>2</sub>0|A7=H<sub>2</sub>0|A8=H<sub>2</sub>0
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|A9=H<sub>2</sub>0|A10=H<sub>2</sub>0|A11=H<sub>2</sub>0|A12=H<sub>2</sub>0
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|B1=H<sub>2</sub>0
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|B2=LB+0µM DAP|B3=LB+20µM DAP|B4=LB+25µM DAP|B5=LB+30µM DAP|B6=LB+35µM DAP
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|B7=LB+40µM DAP|B8=LB+45µM DAP|B9=LB+50µM DAP|B10=LB+55µM DAP|B11=LB+60µM DAP
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|B12=H<sub>2</sub>0|C1=H<sub>2</sub>0
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|C2=S0.2+0µM DAP|C3=S0.2+5µM DAP|C4=S0.2+10µM DAP|C5=S0.2+15µM DAP|C6=S0.2+20µM DAP
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|C7=S0.2+25µM DAP|C8=S0.2+30µM DAP|C9=S0.2+35µM DAP|C10=S0.2+40µM DAP|C11=S0.2+45µM DAP
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|C12=H<sub>2</sub>0|D1=H<sub>2</sub>0
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|D2=S0.4+0µM DAP|D3=S0.4+5µM DAP|D4=S0.4+10µM DAP|D5=S0.4+15µM DAP|D6=S0.4+20µM DAP
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|D7=S0.4+25µM DAP|D8=S0.4+30µM DAP|D9=S0.4+35µM DAP|D10=S0.4+40µM DAP|D11=S0.4+45µM DAP
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|D12=H<sub>2</sub>0|E1=H<sub>2</sub>0
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|E2=S0.6+0µM DAP|E3=S0.6+2µM DAP|E4=S0.6+5µM DAP|E5=S0.6+8µM DAP|E6=S0.6+11µM DAP
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|E7=S0.6+14µM DAP|E8=S0.6+17µM DAP|E9=S0.6+20µM DAP|E10=S0.6+23µM DAP|E11=S0.6+26µM DAP
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|E12=H<sub>2</sub>0|F1=H<sub>2</sub>0
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|F2=S0.6+0µM DAP|F3=S0.6+2µM DAP|F4=S0.6+5µM DAP|F5=S0.6+8µM DAP|F6=S0.6+11µM DAP
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|F7=S0.6+14µM DAP|F8=S0.6+17µM DAP|F9=S0.6+20µM DAP|F10=S0.6+23µM DAP|F11=S0.6+26µM DAP
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|F12=H<sub>2</sub>0|G1=H<sub>2</sub>0
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|G2=S0.8+0µM DAP|G3=S0.8+2µM DAP|G4=S0.8+5µM DAP|G5=S0.8+8µM DAP|G6=S0.8+11µM DAP
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|G7=S0.8+14µM DAP|G8=S0.8+17µM DAP|G9=S0.8+20µM DAP|G10=S0.8+23µM DAP|G11=S0.8+26µM DAP
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|G12=H<sub>2</sub>0|H1=H<sub>2</sub>0|H2=H<sub>2</sub>0|H3=H<sub>2</sub>0|H4=H<sub>2</sub>0
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|H5=H<sub>2</sub>0|H6=H<sub>2</sub>0|H7=H<sub>2</sub>0|H8=H<sub>2</sub>0
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|H9=H<sub>2</sub>0|H10=H<sub>2</sub>0|H11=H<sub>2</sub>0|H12=H<sub>2</sub>0}}

Revision as of 13:33, 24 July 2007

Minipreps

BBa_E0422 in pSB1A2 - Clone 1&2 - MP6.1' & MP6.2'
BBa_E0241 in pSB1A2 - Clone 1 - MP7.1'
BBa_J61047 in pSB1A2 - Clone 1&2 - MP9.1' & MP9.2'

Growth kinetics of w121 strain

We make an array to test growth of w121 on different growth media (LB, S0.2, S0.4, S0.6, S0.8), supplemented with different amount of DAP.

Two questions are addressed by the following assay:

-What is the growth behaviour of w121 (dapA- strain) at different concentrations of DAP? -How does w121 strain grow on filtrates of MG1655 growth medium; that is, does MG1655 secrete DAP during growth?

MG1655 was grown on LB medium and the growth medium was filtered free of bacteria at different DO (Optical Densities) during exponential growth phase:

S0.2 (at DO=0.2) S0.4 (at DO=0.4) S0.6 S0.8


W121 was grown on different media & Growth kinetics were measured:


LB line B in the array S0.2 (at DO=0.2) line C in the array S0.4 line D in the array S0.6 line E & F in the array S0.8 line G in the array

In the different columns, DAP was added to the indicated final concentrations (without taking into account DAP produced by MG1655 regarding the recycled growth media)


Kinetic Array :w121 kinetic as a function of DAP and supplemented medium (S0.x)
1 2 3 4 5 6 7 8 9 10 11 12
A H20 H20 H20 H20 H20 H20 H20 H20 H20 H20 H20 H20
B H20 LB+0µM DAP LB+20µM DAP LB+25µM DAP LB+30µM DAP LB+35µM DAP LB+40µM DAP LB+45µM DAP LB+50µM DAP LB+55µM DAP LB+60µM DAP H20
C H20 S0.2+0µM DAP S0.2+5µM DAP S0.2+10µM DAP S0.2+15µM DAP S0.2+20µM DAP S0.2+25µM DAP S0.2+30µM DAP S0.2+35µM DAP S0.2+40µM DAP S0.2+45µM DAP H20
D H20 S0.4+0µM DAP S0.4+5µM DAP S0.4+10µM DAP S0.4+15µM DAP S0.4+20µM DAP S0.4+25µM DAP S0.4+30µM DAP S0.4+35µM DAP S0.4+40µM DAP S0.4+45µM DAP H20
E H20 S0.6+0µM DAP S0.6+2µM DAP S0.6+5µM DAP S0.6+8µM DAP S0.6+11µM DAP S0.6+14µM DAP S0.6+17µM DAP S0.6+20µM DAP S0.6+23µM DAP S0.6+26µM DAP H20
F H20 S0.6+0µM DAP S0.6+2µM DAP S0.6+5µM DAP S0.6+8µM DAP S0.6+11µM DAP S0.6+14µM DAP S0.6+17µM DAP S0.6+20µM DAP S0.6+23µM DAP S0.6+26µM DAP H20
G H20 S0.8+0µM DAP S0.8+2µM DAP S0.8+5µM DAP S0.8+8µM DAP S0.8+11µM DAP S0.8+14µM DAP S0.8+17µM DAP S0.8+20µM DAP S0.8+23µM DAP S0.8+26µM DAP H20
H H20 H20 H20 H20 H20 H20 H20 H20 H20 H20 H20 H20
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