Media_Preparation - PhytoTechnology Laboratories, Inc....

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PLANT TISSUE CULTURE MEDIA PREPARATION F ormula tion charts posted on the "Technical Literature" page on web site FROM PACKED POWDER Powdered media are extremely hygroscopic and must be protected from atmospheric moisture. If possible the entire contents of each package should be used immediately after opening. Preparing the medium in a concentrated form is not recommended as some salt added to the medium may affect shelf life and storage conditions. The basic steps for preparing the culture medium are listed below: 1. Measure out approximately 90% of the final required volume of tissue culture grade water, e.g. 900 ml for a final volume of 1000 ml. Select a container twice the size of the final volume. 2. While stirring the water add the powdered medium and stir until completely dissolved. 3. Rinse the original container with a small volume of tissue culture grade water to remove traces of the powder. Add to the solution in Step 2. 4. Add desired heat stable supplements (e.g. sucrose, gelling agent, vitamins, auxins, cytokinins, etc.) 5. Add additional tissue culture grade water to bring the medium to the final volume. 6. While stirring, adjust medium to desired pH using NaOH, HCl, or KOH. 7. If a gelling agent is used, heat until the solution is clear. 8. Dispense the medium into the culture vessels before (or after) autoclaving according to your application. Add heat labile constituents after autoclaving. 9. Sterilize the medium in a validated autoclave at 1 kg/cm 2 (15 psi), 121 ° C, for the time period described under Sterilization of Media. 10. Allow medium to cool prior to use. *Heating may be required to bring powders into solution. POWDERED MEDIA AND BASAL SALT MIXTURES ARE FOR LABORATORY USE ONLY.
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This note was uploaded on 07/23/2011 for the course HOS 6737c taught by Professor Moore during the Spring '09 term at University of Florida.

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Media_Preparation - PhytoTechnology Laboratories, Inc....

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