I want to do MIC for a crude extract, kindly provide me with the procedure for MIC.
regards
Nuzhath
Dear Nuzhath, you can follow CLSI or EUCAST manuals for determination of MIC values for antibiotics. If you have plant extract, you can follow similar question and answers in the link indicated below.
https://www.researchgate.net/post/I_am_not_able_analyze_the_pattern_of_zone_for_the_disc_diffusion_assay_of_my_plant_extract
Hi
By following the instruction of CLSI you can do your MIC for antobiotics
regards
Hi,
Here is the protocol I use for MIC determination of Staphylococcus.
Staphylococci were grown with aeration in 50-ml falcon tube containing 5 mL Tryptic Soy Broth (TSB; Becton Dickinson and Company, Franklin Lakes NJ, USA) in a shaker maintained at 37˚C. The medium was supplemented with 6 µg/ml vancomycin as required. The overnight culture was diluted with cation-adjusted Muller–Hinton Broth (MHB; Becton Dickinson and Company) to have approximately 5 x 104 colony forming units (CFU) per 100 µl in each well in a round bottom 96-well plate. Serial dilutions of antibiotics were added to each well to obtain a final concentration of 128 µg/ml to 0.0125 µg/ml. The plates were further incubated at 37˚C for 20 h and MIC was determined as the lowest concentration that did not allow visible growth.
5x104 CFU per well is about 100 fold dilution after maintaining the OD600 to 0.5
Regards,
Suresh
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