| Component | M.W. | mM | amount/100 ml | amount/500 ml |
| 1. CaCl2.2H2O | 147.02 | 2.0 | 30.0 mg | 150 mg |
| 2. KCl | 74.56 | 3.2 | 23.5 mg | 118 mg |
| 3. MgCl2.6H2O | 203.3 | 0.5 | 10 mg | 50 mg |
| 4. NaH2PO4.H2O | 137.99 | 0.4 | 5.6 mg | 28 mg |
| 5. Lactic Acid (Sodium salt; 60% w/w syrup) | 112.1 | 10.0 | 186
mg (141 µl) |
930
mg (705 µl) |
| 6. NaCl | 58.44 | 114.0 | 666 mg | 3330 mg |
| 7. NaHCO3 | 84.01 | 2.0 | 16.8 mg | 84 mg |
| 8. HEPES (Free Acid) | 238.3 | 10.0 | 238 mg | 1190 mg |
| 9. Na Pyruvate (from stock) | 110.0 | 0.2 | 1.0 ml | 5.0 ml |
| 10. BSA (Fraction V) | ------ | 3 mg/ml | 300 mg | 1500 mg |
| 11. Gentamycin (from stock) | ------ | 50 µg/ml | 100 µl | 500 µl |
| Add approximately 80% of the double distilled water need to a beaker. With stirring, add components 1 - 8 in order. Adjust pH to 7.4 at 20-22 C using 1 or 2 N NaOH. Add water to bring to 100 % of volume. This can be stored up to two weeks as a stock at 5 C if filtered and kept sterile. Generally we make up 500 ml at a time. On day of use, add components 9 and 10. Note that these can be for any volume desired for use that day and need not be 100 or 500 ml. Filter to sterilize with 0.2 µm filters. See Stock solutions for components 9 and 11. |