I conducted experiments of cup diffusion agar method I noticed the extract diffusion differed according to plant and solvent of extraction, Therefore, I agree with the answers that recommend to make different times separately
Good luck
24 hrs enough for optimum time but according to some articles show that it may be 3 hours in some plants.
If the optimum is unknown, do the necessary experiments to determine the optimum.
I use to use overnight at 4C (13 to 16 hours).
That being said, I like Dr Klvana's answer too.
Also, I use to put the antibiotics in the agar and mix with the agar just as the plates are solidifying, and being poured, so diffusion was not needed.
Many times I have reheated my 'plain' agar back up..to make liquid again.
I use to keep an old beaker with a stir-bar in it in the 4C with like half full of agar.
When I needed plates I would place him on the hot-plat and in a few minutes
had my plate, and would add my antibiotic(s) when the agar cooled just before casting.
Best Wishes!
Dear Mac,
I did not try my own work. But i saw it in an article. I think you can try 3 hours, 12 hours and 24 hours separately. if you have extra material. Thus, interval times are also determined. this is just İn my opinion :)
May be 3 hours is the optimum.but i advise you to try the three times separately .
I think it depends on the nature of the diffusion of the plant extract. Some plant extracts are diffusing rapidly but the other require much time to do. To ensure that all your plant extracts are diffused through the agar plate , you have to incubate them for 24 hours to obtain excellent results of complete diffusion.
I think it depend on the active ingredient of plant extraction and it diffusion in plate agar and another factor is concentration of active ingredient of the plant.
finally I think it's depend on plant under experiment
I conducted experiments of cup diffusion agar method I noticed the extract diffusion differed according to plant and solvent of extraction, Therefore, I agree with the answers that recommend to make different times separately
Good luck
In fact, I relied in my researches on a 24-hour incubation that gives more favorable results for spreading plant extracts, you can use different times of incubation to determine the highest results of the inhibition zone.
Agar pre-diffusion at room temperature or at 4°C? 4°C would be better, for two hours followed by an incubation at the ideal temperature for the target microbe.
Ithink it depends on the nature of the difussion of the plant extract 24 hours excellant .
Dear dr.
This depends on the nature of the research and the bacteria on which the examination is performed. For example, if the examined bacteria were food born bacteria , then the effect of the extracts as preservative on the bacteria should be studied in 4 ° C conditions, as well as incubation at room temperature, compared to standard conditions.
for two hours followed by an incubation at the ideal temperature for the target microorganism
Article Methods for in vitro evaluating antimicrobial activity: A review
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