Orchid Maintenance/ Replate Medium

COMPOSITION

INSTRUCTION FOR USE

Dissolve 33.36gms of dehydrated medium in 600ml of distilled or deionized water at room temperature (15-30^oC). 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. Sometimes 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 1000ml with distilled water. Mix gently, heat and rotate between intervals until the solution becomes clear. Do not boil, reheat and allow to cool below 50^oC during dispensing. Dispense the medium into suitable containers, plug or cap, then autoclave at 15psi (121^oC) 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 45-500C and aseptically add desired sterile heat-labile 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.

STORAGE

Dehydrated plant tissue culture media is hygroscopic and should be protected from sunlight and moisture. Store the prepared medium at 2–8°C away from direct light. Medium should be used before the expiry date. The entire volume of each bottle should be used immediately after opening or else the unused portion should be stored in desiccators and refrigerated at 2–8°C.