Directions for the preparation of single strength synthetic liquid media (1X) from concentrated media
- Measure out sterile culture-grade water (Cat. # 03-055-1) to approximately 70% of desired total volume of medium. Pour water into an appropriate sterile mixing container that is close to the desired final volume. The water should be at room temperature.
- Add the amount of the concentrated medium.
- Add the desired amount of L-Glutamine Solution 200mM (Cat. # 03-020-1) if required (See table)*.
- Add the desired amount of Sodium Bicarbonate Solution (Cat. # 03-040-1). (See table).
- Add antibiotics solution if desired*.
- Add water to the final volume. During the dilution, mix gently to obtain a uniform solution. If necessary, adjust pH with sterile 1N NaOH or HCl.
- Store at 2ºC to 8ºC.
* To avoid shortening the shelf life, add these components just before use.
Important: The above procedures are carried out under strict sterile environment.
Example I, without sterile filatration (asceptic technique)
Preparation of Basal Medium Eagle, Earle’s Salts Base, one liter:
- 700 ml sterile water (Cat. # 03-055-1)
- 100 ml Basal Medium Eagle (Earle's) 10X w/o L-Glutamine, w/o Sodium Bicarbonate (Cat. # 01-015-5).
- 10 ml L-Glutamine Solution 200mM (Cat. # 03-020-1)
- 29.4 ml Sodium Bicarbonate Solution 7.5% (Cat. # 03-040-1).
- 10 ml Penicillin-Streptomycin Solution (Cat. # 03-031-1)
- Adjust pH if necessary with sterile 1N NaOH or 1N HCl. Sterile water to final volume.
- Example II, with sterile filtration (non-asceptic technique)
Example II, with sterile filtration (non-asceptic technique)
Preparation of RPMI 1640, 1 liter:
- 700 ml distilled water
- Add 100 ml RPMI concentrate 10X
- Adjust pH to 6.5-7.0 with 1N NaOH
- Add 10.3 ml L-Glutamine Solution 200mM
- Add 26.7 ml Sodium Bicarbonate Solution 7.5%
- Adjust pH with 1N NaOH or 1N HCl to pH 7.0-7.4
- Add distilled water to final volume. Adjust pH if necessary.
- Filter the medium into sterile container using a 0.2µm membrane filter.
Recommended amounts of Sodium Bicarbonate and L-Glutamine to be added in the preparation of single strength liquid media (1X) from concentrated media (5X or 10X)
| Product Description | Desired Cat. No. | Prepared from Cat. No. | Sodium Bicarbonate Solution 7.5%, | L-Glutamine Solution 200mM, |
|---|---|---|---|---|
| Cat.# 03-040-1 | Cat.# 03-020-1 | |||
| ml/Liter | ml/liter | |||
| Basal Medium-Eagle Earles salts base | 01-015-1 | 01-015-5 | 29.4 | 10 |
| Minimum Essential Medium Eagle Earles Salts Base | 01-025-1 | 01-025-5 | 29.4 | 10 |
| Dulbeccos Modified Eagle Medium (DMEM), Low Glucose | 01-050-1 | 01-050-4 (5X) | 49.4 | 20 |
| Dulbeccos Modified Eagle Medium (DMEM), High Glucose | 01-055-1 | 01-055-4 (5X) | 49.4 | 20 |
| Glasgow Modified MEM*(GMEM) | 01-060-1 | 01-060-5 | 36.7 | 20 |
| Medium M-199 Earles Salts Base | 01-080-1 | 01-080-5 | 29.4 | |
| Medium M-199 Hanks Salts Base | 01-085-1 | 01-085-5 | 4.7 | |
| Nutrient Mixture F10 (HAM) | 01-090-1 | 01-090-5 | 16 | |
| Nutrient Mixture F12 (HAM) | 01-095-1 | 01-095-5 | 15.7 | |
| RPMI 1640 | 01-100-1 | 01-104-5 | 26.7 | |
| 10.3 | ||||
| DMEM F12 (1:1) | 01-170-1 | 01-170-5 | 16 | |
| 12.5 | ||||
| DCCM-1 | 05-010-1 | 05-010-5 | 29.4 | 10-20 |
| DCCM-2 | 05-015-1 | 05-015-5 | 29.4 | 10-20 |
| LPM | 05-040-1 | 05-040-5 | 29.4 | 10-20 |
| BIO-MPM-1 | 05-060-1 | 05-060-5 | 26.9 | 10-20 |
| BIOCHO-1 SFM Base | 05-061-1 | 05-061-5 | 26.9 | 10-20 |
| BIOCHO-2 SFM Base | 05-062-1 | 05-062-5 | 4.7 | 10-20 |
