Growth curve measurement

Reagents:

1. Staphylococcus aureus solution (37℃, 200rpm, overnight culture);
2. Anti-microbial peptides stock solutions (1mg/ml) (or AMPs: LL-37, Grammistin-Pp1, GF-17) (China peptides, Suzhou)
3. LB Broth.

Procedure:

1. Add 100ml LB Broth into each 250ml conical flask (5 in total);
2. Label each of conical flask as LL-37, Grammistin-Pp1, GF-17, Mix, and Staphylococcus aureus.
3. Add 300μl Staphylococcus aureus solution (OD₆₀₀≈4) into each conical flask.
4. Culture at 37℃, 180rpm.
5. Measure OD₆₀₀ value after each hour, when OD₆₀₀ value is 0.05, add 300μl anti-microbial peptides into each conical flask except the one labeled Staphylococcus aureus (the concentration of peptides is 1mg/ml). Measure OD₆₀₀ value every 2 hour until the Staphylococcus aureus grow into the stationary phase (12 hours)