W/Calcium Chloride, Vitamins, Sucrose and Agar
Instructions for use - Dissolve 41.54g of dehydrated medium in 600 ml of distilled or deionized water at room temperature (15-30C). Rinse media vial with small quantity of distilled water to remove traces of powder. Add the desired heat stable supplements prior to autoclaving. Continue stirring until the powder has dissolved. Some times media does not dissolve completely unless the pH is reduced. For these, lower the pH to about 3.0 to facilitate dissolution of media. The pH of medium is adjusted by using 1N HCL/ 1N NaOH/ 1N KOH. Make up the final volume to 1000 ml with distilled water. Mix gently, heat and rotate between intervals until the solution becomes clear. Do not boil, reheat and allow to cool below 50C during dispensing. Dispense the medium into suitable containers, plug or cap, then autoclave at 15psi (121C) for 15 minutes, using a slow exhaust cycle. Higher temperatures and/or longer times are not recommended. Cool the autoclaved culture vessels containing medium to 45-50C and aseptically add desired sterile heatlabile substrate. Note: Media should be prepared according to formula mentioned on the label however, it is recommended to use an entire container at once. Heat-labile substrates should be added, after autoclaving.