Paris/July 7
From 2007.igem.org
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 amounts 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 etc S0.8
W121 was grown on different media:
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 in the array S0.8 line F 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 | |
| B | H20 | LB+0µM DAP | LB+25µM DAP | LB+37.5µM DAP | LB+50µM DAP | LB+62.5µM DAP | LB+75µM DAP | LB+100µM DAP | LB+300µM DAP | LB+0µM DAP | H20 | |
| C | H20 | S0.2+0µM DAP | S0.2+25µM DAP | S0.2+37.5µM DAP | S0.2+50µM DAP | S0.2+62.5µM DAP | S0.2+75µM DAP | S0.2+100µM DAP | S0.2+300µM DAP | S0.2+0µM DAP | H20 | |
| D | H20 | S0.4+0µM DAP | S0.4+25µM DAP | S0.4+37.5µM DAP | S0.4+50µM DAP | S0.4+62.5µM DAP | S0.4+75µM DAP | S0.4+100µM DAP | S0.4+300µM DAP | S0.4+0µM DAP | H20 | |
| E | H20 | S0.6+0µM DAP | S0.6+25µM DAP | S0.6+37.5µM DAP | S0.6+50µM DAP | S0.6+62.5µM DAP | S0.6+75µM DAP | S0.6+100µM DAP | S0.6+300µM DAP | S0.6+0µM DAP | H20 | |
| F | H20 | S0.8+0µM DAP | S0.8+25µM DAP | S0.8+37.5µM DAP | S0.8+50µM DAP | S0.8+62.5µM DAP | S0.8+75µM DAP | S0.8+100µM DAP | S0.8+300µM DAP | S0.8+0µM DAP | H20 | |
| G | H20 | H20 | H20 | H20 | H20 | H20 | H20 | H20 | H20 | H20 | H20 | |
| H | ||||||||||||
See results.
The 96 well plate is surrounded by H2O in order to maintain humidity during the assay. In each slot, 200µL of the growth medium (LB or S0.x) is mixed with 2µL of w121 culture grown ON, and with different amount of DAP (see table). The growth profile is measured over a period of 10H, a DO measurement is acquired each 3min.
