Murashige and Skoog Medium, without Sucrose, Agar, IAA, and (C08-0503-902)

Chemicals are carefully selected and, whenever possible, analytical grade materials were used. Finished batches are checked to ensure that they conform to preset high standards before they are released for sale. Except where otherwise indicated, the amounts shown relate to single strength media. Preparation of Liquids from Powder Plant Tissue Culture Media: 1. Measure out approximately 25% less purified water (distilled or deionized) than the final volume of medium required. The water should be at room temperature (15°C to 30°C). 2. Stir the water and slowly add the powder. Rinse out the inside of the container to remove all traces of powder. Continue stirring until the powder has dissolved. Some media will not dissolve completely unless the pH is reduced. For these, lower the pH to about 3.0 to facilitate solution (Note: It is possible to weigh out the desired quantity of powder from a container, using the weight noted on the product label. It is recommended, however, to use an entire container at once, in which case it is not necessary to weigh the powder). 3. Add the required supplements and stir to disperse (Note: Heat-labile substrates should be added at step 8, after autoclaving). 4. Check and if necessary, adjust the pH of the medium to the desired level (normally 5.5 + 0.1). Mix gently during additions. 5. Add sufficient purified water to give a volume equal to the final volume less any heat-labile substance to be added in step 8. 6. Add the desired quantity to agar. Heat, with continuous mixing, until the solution is clear. Do not boil. Do not allow to cool below 50°C during dispensing. 7. Dispense the medium into suitable containers, plug or cap, then autoclave at 121°C (1 bar, 15 psi) for 15 minutes, using a slow exhaust cycle. Higher temperatures and/or longer times are not recommended. 8. After cooling, aseptically add desired sterile heat-labile supplements. 9. Label and store at 2°C to 8°C. Murashige and Skoog Medium is widely used plant tissue culture growth medium. It has proven effective in the culture of tissue derived from moncotyledons and dicotyledons. This medium, originally formulated to support tobacco callus, has been used to support callus initiation and growth, the growth of cells in suspension culture and the regeneration of shoots and plantlets from explants. More recently, it has been used for micropropagation of ornamental, vegetable and fruit plants in research and commercial enterprises. Murashige and Skoog Medium contains Murashige and Skoog salts, as well as a mixture of vitamins. This modification of the original formula does not contain agar, sucrose, IAA, and Kinetin.