All powdered cell culture media are with L-Glutamine and without sodium bicarbonate.
Procedure:
- To a mixing container that is as close to the final volume as possible, add 10% less distilled water than the desired total volume of medium.
- Add powdered medium to room temperature water with gentle stirring. Do not heat water.
- Rinse inside of package to remove all trace of powder.
- Add Sodium Bicarbonate as required.
- Dilute the medium to the desired volume with distilled water and stir until dissolved. Do not overmix.
- Adjust the pH to between 0.2 – 0.3 below the desired final working pH by slowly adding, with stirring, 1NaOH/ 1N HCI. The pH usually will rise 0.2 – 0.3 units upon filtration. Keep the container closed until the medium is filtered.
- Process the medium immediately into sterile containers by membrane filtration using 0.2 micron filter.
Example I
DMEM (11-055-1) – 1 liter
- Prepare 900ml of distilled water in clean glass beaker. Water temperature should be 15-30ºC. Put the beaker on a stirrer and add a magnet.
- Add the 1-liter powder to the water and stir gently. Fill some distilled water into the empty package, stir and pour the remains into the beaker. Stir till completely dissolved.
- Add 3.7 gram Sodium Bicarbonate (or 49.4 ml of 7.5% Sodium Bicarbonate Solution).
- Adjust pH to 0.2-0.3 units below the required pH using 1N HCl or 1N NaOH. The pH will rise by 0.1-0.3 units after filtration.
- Add distilled water up to 1 liter.
- Filter for sterility with 0.2µ filter into sterile bottles.
- For the preparation of 10 liters multiply by 10.
Example II
RPMI 1640 (11-100-1) – 1 liter
- Prepare 900ml of distilled water in clean glass beaker. Water temperature should be 15-30ºC. Put the beaker on a stirrer and add a magnet.
- Add the 1-liter powder to the water and stir gently. Fill some distilled water into the empty package, stir and pour the remains into the beaker. Stir till completely dissolved.
- Add 2 gram Sodium Bicarbonate (or 26.67 ml of 7.5% Sodium Bicarbonate Solution).
- Adjust pH to 0.2-0.3 units below the required pH using 1N HCl or 1N NaOH. The pH will rise by 0.1-0.3 units after filtration.
- Add distilled water up to 1 liter.
- Filter for sterility with 0.2µ filter into sterile bottles.
- For the preparation of 10 liters multiply by 10.
Sodium Bicarbonate Concentrations:
| Cat. No. | gram/liter | ml/liter from 7.5% Solution |
| 11-025-1 | 2.2 | 29.3 |
| 11-040-1 | 2.2 | 29.3 |
| 11-042-1 | 2.2 | 29.3 |
| 11-050-1 | 3.7 | 49.3 |
| 11-055-1 | 3.7 | 49.3 |
| 11-058-1 | 3.024 | 40.32 |
| 11-060-1 | 2.75 | 36.67 |
| 11-075-1 | 2.2 | 29.3 |
| 11-080-1 | 2.2 | 29.3 |
| 11-085-1 | 0.35 | 4.67 |
| 11-090-1 | 1.2 | 16 |
| 11-095-1 | 1.176 | 15.68 |
| 11-100-1 | 2 | 26.67 |
| 11-115-1 | - | - |
| 11-170-1 | 1.2 | 16 |
