Yes, this product in solution can be autoclaved for sterilization. Please find the preparation instructions below.
This powder is extremely hygroscopic and must be protected from atmospheric moisture. Do not open the container until its contents are allowed to warm to room temperature. If possible the entire contents of the package should be used immediately after opening.
1.6 g of powder is required to prepare 1 L of medium.
Preparing this product in a concentrated form is not recommended as some salt complexes may precipitate. Supplements may be added prior to sterilization or added aseptically to a sterile medium. Certain supplements (i.e. heat labile) may require filter sterilization and may affect the shelf life of the medium.
The basic steps for preparing culture medium are the following:
1. Using a container twice the size of the desired final volume, measure out approximately 90% of the final required volume of tissue culture grade water (e.g. Sigma Product No. W-3500). Example: 900 ml for a final volume of 1000 ml.
2. While stirring, add the powdered product.
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 any desired supplements (e.g. vitamins, gelling agent, auxin, cytokinin, etc.).
5. While stirring, adjust to the desired pH (e.g. 5.7 +/- 0.1) using KOH, NaOH, or HCl.
6. Add additional tissue culture-grade water to bring the medium to the final volume.
7. If a gelling agent is used, heat the solution to approximately 100°C while stirring.
8. Dispense the medium into 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/cm2 (15 psi). The medium should attain a temperature of 121°C for at least 15 min.
All media preparations should be stored at 0-5°C. Store dry powder in a desiccator. Deterioration of powdered medium may be recognized by the following: 1) color change; 2) granulation, clumping, or particulate matter throughout the powder; 3) pH change; and 4) inability to promote growth when properly used.
