"

From 2010.igem.org

Performing a Kill Assay

-Making 4 different catechol concentrations: 0mM, 0.5mM, 1mM, and 2.5mM

-Set the shaker to 30C

-Making a catechol stock solution-->0.05M

Analytical balance used

    0.0275g = 5mL
    actual mass = x           
    x--> how much water is needed

Calculation:

  0.0275g = 5mL
  0.0118g = x
  x=2.145mL

-Diluted ovn from 1.281 OD600 to 0.958

-Added 500uL into the corresponding (*) tubes with their respective amount of LB

-Added corresponding amounts of stock catechol solution to each tube(*)

(*)-->see protocol for corresponding amounts

-Plated 10uL of each solution → incubated at 37C for 2 hours (11:30am-1:30pm)

-Put the remaining solutions into the shaker at 30C for 2 hours (11:30am-1:30pm)

-->the temp fluctuated and was a bit over 30C

2hr mark (1:30pm)

-->plated the 4 solutions onto the “2hr” plate (0 and 0.5 are switched)

4hr mark (3:30pm)

-->plated in 4hr plate (0mM, 0.5mM, 1mM, and 2.5mM)