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Reconstitute the contents of one vial of 2 mg Mycobactine J with 2 ml absolute ethanol (analytical grade). Shake carefully so that the product is completely dissolved. Pour the contents of the bottle into 1 litre of medium 645 kept melted in a water bath at +56°C, taking care to pour out all of the product. Rinse out the vial that contained the Mycobactine J with an additional 2 ml of absolute ethanol and pour into the culture medium. Mix the Mycobactine J alcohol solution and the culture medium very thoroughly.
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