Protocol for: Minimum inhibitory concentration (MIC)
Abstract or Description:
Minimum Inhibitory Concentration
1. Grow cultures up overnight.
2. The following day, inoculate a fresh culture with a 1:10-20 dilution and grow up to OD600 0.4.
3. Dilute to OD600 0.0005 (1:800) in THB (or relevant media).
4. Dilute this 1:200 and drop about 25µl on THA (or relevant agar plates) to confirm equal starting inoculums.
5. Put 50µl of the 0.0005 cultures in a 96 well plate in triplicates.
6. Add 50µl per well of the substance you want to test (ie. antimicrobial peptide, antibiotic) to each well, keeping in mind the final concentration will be half the original concentration you are adding. It is good to test 1:2 dilutions (in THB, or relevant media) of this substance.
7. Incubate overnight and check by eye or OD600 24 hours later to determine what concentration inhibited the growth of the different strains of bacteria.
Posted By: A Timmer
on 1/12/2009 6:26:42 PM