I need some suggestions on working with the drug 5-fluoroindole. I am working on green microalgae and trying to make my strain tryptophan auxotroph. However, I am unable to make proper drug plates as the desired drug concentration is extremely low, hence difficult to measure. Secondly, 5-FI being light sensitive, it takes longer for the colonies to be visible if there are any since I need to keep the plates away from light.
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Rana Hadi Al-Shammari
Preparation of 5-Fluoroindole Stock Solution:Utilize amber glassware or aluminum foil to shield the drug from light exposure during preparation. Dissolve 5-fluoroindole in a suitable solvent known to enhance solubility (e.g., dimethyl sulfoxide, DMSO) at a concentration exceeding its solubility limit. Employ gentle heating or sonication if necessary to facilitate dissolution. Exercise caution to prevent excessive exposure to heat, which may degrade the compound. Filter the solution through a sterile filter (e.g., 0.2 μm pore size) to remove particulate matter and ensure sterility. Store the stock solution in opaque vials or wrapped in aluminum foil at a temperature conducive to stability (-20°C is recommended). Preparation of Agar Medium:Select a suitable agar-based medium compatible with the growth requirements of the target microorganism or experimental system. Common choices include nutrient agar or Sabouraud agar for bacterial and fungal cultures, respectively. Follow standard protocols for the preparation of agar medium, incorporating appropriate nutrients and pH adjustments as per experimental requirements. Ensure that the agar medium is free from contaminants and adequately sterilized through autoclaving or filtration. Incorporation of 5-Fluoroindole into Agar Plates:Under subdued light conditions, aseptically dispense the desired volume of the 5-fluoroindole stock solution into the molten agar medium at a predetermined concentration, considering the desired final concentration and solubility limitations. Thoroughly mix the agar medium to ensure uniform dispersion of 5-fluoroindole throughout the medium while minimizing exposure to ambient light. Pour the agar medium containing 5-fluoroindole into sterile petri dishes, ensuring even distribution and adequate solidification. Seal the plates with parafilm or aluminum foil to further shield the drug from light.
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