I've already run the test and all tubes give turbidity, but I can't know whether it's due to the compound precipitate or to bacterial growth.
I've read adding resazurin (Alamar blue) or a tetrazolium salt to the tubes would give a change in colour in case of bacterial growth. If this is the case, would this method tell me the MIC or the MBC?
MIC by broth dilution method in 96 well titre plate
for growth indicator you can add INT, which give pink color if growth is present in dilution.
I've used test tubes, therefore it's broth macrodilution. Is there any difference?
Btw, what quantity and in which state (solid, aqueous dilution) should I add the INT?
Thank you.
If your compound precipitates or is insoluble at higher concentrations, you won't get an accurate MIC value. When I work with compounds that have a poor solubility in water, I make a highly concentrated solution in DMSO and dilute this into broth to make my starting concentration. Then do serial dilutions as you would normally. You need to do a control with that DMSO concentration in broth to make sure it's not affecting your cells. 3% and lower is fine but above this you start to see decreased cell growth relative to no DMSO.
First you dilute your sample in DMSO in higher concentation then two fold dilute the sample in DMSO only then you add in broth
please read this article might you get answer
http://www.ijpsonline.com/article.asp?issn=0250-474X;year=2014;volume=76;issue=2;spage=148;epage=156;aulast=Rakholiya
You can try by Well agar diffusion methods, and try to consult this reference:
Pridham TG, Lindenfelser LA, Shotwell OL, Stodola F, Benedict
RG, Foley C, Jacks PW, Zaumeyer WJ, Perston WH,
Mitchell JW. Antibiotics against plant disease. I. Laboratory
and green house survey. Phytopathol 1956; 46: 568–575.
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