as I am working first time on antibiotics for analysis of MIC by broth dilution method.
I firstly prepared stock soln of 100mg/ml of ampicillin.
now I want to ask how much ampicillin from its stock solution I should transfer to 100microliter of LB medium in 96 microtiter plate ?
In order to determine the MIC you will need to do a range of concentrations. Probably from 1ug/ml up to 100ug/ml or more. As to how much to add, it is just simple math. You have 100mg/ml to begin with, so a 10-fold dilution would be 10mg/ml and 100-fold dilution would be 1mg/ml etc. So if you have 100ul of medium, then 1/10 of that volume (10ul) would be 10mg/ml, and 1/100 of that volume (1ul) would be 1mg/ml.
According to the CLSI guide line, 256 mg/L is the highest concentration. You should experiment by two fold dilution statr with this concentration
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