Lyophilized peptides were solubilized in sterile Milli-Q water to a final concentration of 1 mM and filter sterilized through a 0.22 m pore filter. Minimum inhibitory concentration (MIC) of the peptides was assessed as previously described [11] using a growth inhibition assay. Dilutions of the peptides were made to obtain a final concentration of 100, 50, 25, 12.5, 6.2, 3.1, and 1.6 μM, and in the case of the peptide mixture, an equimolar solution was used. Kanamycin sulfate (Sigma, St. Louis, MO, USA) was included for E. amylovora as a reference antibiotic. Twenty microlitres of each dilution were mixed in a microtiter plate well with 20 μL of the corresponding bacterial suspension (final concentration 107 CFU mL−1) and 160 μL of Tryptic Soy Broth (TSB) (BioMèrieux, Marcy-l’Étoile, France) to a total volume of 200 μL. Three replicates for each peptide and concentration were used. Bacterial growth was determined by optical density measurement at 600 nm (Bioscreen C, Labsystem, Helsinki, Finland). Microplates were incubated at 25 °C with 20 s shaking before hourly absorbance measurement for 48 h. The experiment was repeated twice. The MIC was taken as the lowest peptide concentration with no growth at the end of the experiment.

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.