by using the overlay method of antagonism the results are good and there is a clear zone but with ethyl acetate extract there is no results.
Liquid media based production and solvent extractions are best choice of methods.
Dr/ Manikandan Madheslu i used tryptic soy broth media and ethyl acetate solvent but the results are negative although the results with cells are positive.
Marwa, try to evaporate the ethyl acetate and reconstitute in aqueous solvent (simple buffer)
All the Best
Regards,
Dr. M. Manikandan
Try with starch casein broth or starch nitrate broth and optimize the inoculum.
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