Hi,

I am conducting a biofilm prevention assay (adding compounds at time of inoculation). Would inoculated broth just be the overnight culture?

Hi,

For biofilm prevention assay, the inoculation broth could be prepared as the following:

1. Prepare 100µl of the fresh medium (e.g. LB) with a desired concentration of the compound into each well of the 96-well plate

2. Dilute the overnight culture (E.coli, P.aeruginose) 1:100 into fresh medium (e.g. LB)

3. Add 10µl of the bacterial dilution into each well of the 96-well plate prepared in step 1. Usually, we replicate 3-4 wells for each treatment for quantitative assays

4. Incubate the plate at 37˚C for 4-24h

After several trials, you can modify the concentration of the compound or the concentration of bacterial culture to achieve an optimal protocol for each specific compound.

For the positive control, you will add only 100µl of the fresh medium (without the compound) in step 1, the remaining steps are the same.

Goodluck.