Toronto/3 August 2010
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)