| Component | M.W. | mM | amount/100 ml |
| 1. CaCl2.2H2O | 147.02 | 2.0 | 29.4 mg |
| 2. KCl | 74.56 | 3.1 | 23.1 mg |
| 3. MgCl2.6H2O | 203.3 | 0.4 | 8.1 mg |
| 4. NaH2PO4.H2O | 137.99 | 0.3 | 4.1 mg |
| 5. Lactic Acid (Sodium salt; 60% w/w syrup) | 112.1 | 21.6 | 368 µl |
| 6. NaCl | 58.44 | 87.0 | 508 mg |
| 7. Na Pyruvate | 110.0 | 1.0 | 11 mg |
| 8. NaHCO3 | 84.01 | 10.0 | 84 mg |
| 9. HEPES (Free Acid) | 238.3 | 40.0 | 953 mg |
| 10. BSA (Fraction V) | ------ | ----- | 600 mg |
| Add approximately 80 ml double distilled water to a beaker. With stiring add components 1 - 9 in order. PH the media to 7.54 at 20-22 C using 5 N NaOH. Add water to bring to 100 ml. This can be stored up to two weeks as a stock at 5 C if filtered and kept sterile. On day of use, add component 10. The pH of the final medium is 7.4 at 39 C. If planning to use at another incubation temperature, the change in the medium pH with temperature is -0.008 pH units / degree C. |