Materials Required
- 1000 ml of Distilled Water
- 4.54 g of MS Media
- 6-10 g of Agar
- 30 g of Sugar
Directions
- Dissolve 4.54 grams of the dry medium in 800 ml of distilled or deionized water at room temperature (20-25*C).
- Rinse the media vial with a small quantity of distilled water to remove traces of power.
- Add the Agar and Sugar.
- Continue stirring until the powder has dissolved.
- Sometimes media does not dissolve completely unless the pH is reduced. So if required, lower the pH to about 3.0 to facilitate the dissolution of media.
- Adjust the pH of the media using 1N HCL, 1N NaOH, or 1N KOH as desired.
- Then add the rest of the water (to make up the volume to 1000 ml or 1 L).
- Add gelling agent (Agar, Gellan Gum, Gelzan, etc.)
- Mix Gently, heat, and rotate between intervals until the solution becomes clear.
- Do not boil, reheat and allow to cool below 50 ℃ during dispensing. Dispense the medium into suitable containers, loosely plug or cap, then autoclave at
1 atm (121 ℃) 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 45-50 ℃ and aseptically add desired sterile heat-labile supplements.
Notes
- Where autoclaving isn’t possible due to lack of equipment it can be replaced with 15 minutes boiling in a pressure cooker for similar levels of sterilisation.
- Alternatively these steps can be replaced with simple boiling for five minutes. This will kill most microbes, but occasionally some types of algae and fungus can survive.
- MS Medium is extremely hygroscopic and will rapidly draw moisture from the air. For this reason it’s important to never leave it exposed to air when not absolutely required.
