The preparation instructions for this product are as follows:
1. Using a container twice the size of the desired final volume, measure out ~90% of the required final volume of tissue culture grade water (e.g., Catalog Number W3500). 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 desired supplements (e.g., sucrose, gelling agent, auxins, cytokinins).
Add heat labile constituents after autoclaving.
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 clarity while stirring.
8. Dispense the medium into culture vessels before or after autoclaving according to the 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 minutes.
Please see the link below to review this and additional information available in the product datasheet:
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/361/537/m5519pis.pdf