Difference between revisions of "Team:XJTLU-CHINA/Protocols Inhibition ring"
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Latest revision as of 08:40, 31 October 2017
Inhibition ring
Reagents:
Staphylococcus aureus solution (37℃, 200rpm, overnight culture); Anti-microbial peptides stock solutions (1mg/ml) (or AMPs: LL-37, Grammistin-Pp1, GF-17) (China Peptides, Suzhou); LB agar plates
Procedure:
- Defreeze AMP solutions at 4℃.
- Add 170μl of LL-37, Grammistin-Pp1, GF-17 solutions to form a 1:1:1 LL-37+Grammistin-Pp1+GF-17 solution.
- Measure OD600 value of Staphylococcus aureus after 9 hours’ culture.
- Inoculate 100μl diluted S. aureus (OD600≈0.4) on each medium.
- Pipette 20μl each peptide solution on each paper disc (LL-37, Grammistin-Pp1, GF-17, and LL-37+Grammistin-Pp1+GF-17).
- Dry all discs.
- Place 2 paper discs with peptide, 1 paper disc with positive control (chloramphenicol) and 1 negative control (buffer: PBS+BSA+HAC) for each plate.
- Overnight culture at 37℃ condition.
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