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)

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.