To prepare L Modified Barth's Saline 10×
Table 1. Recipe
Name | Amount | Concentration | Formula/M.W. | CAS # |
---|---|---|---|---|
Sodium chloride | 46.752 g | 0.8 M | NaCl/58.44 | 7647-14-5 |
Potassium Chloride | 0.745 g | 0.01 M | KCl/74.55 | 7447-40-7 |
2.464 g | 0.01 M | MgSO4·7H2O/246.47 | 10034-99-8 | |
HEPES(N-2-Hydroxyethylpiperazine-N'-2-ethanesulfonic acid) | 11.915 g | 0.05 M | C8H18N2O4S/238.3 | 7365-45-9 |
Sodium bicarbonate | 2.1 g | 0.025 M | NaHCO3/84.01 | 144-55-8 |
- Prepare 0.8 L of distilled water in a suitable container.
- Add 46.752 g of Sodium chloride to the solution.
- Add 0.745 g of Potassium Chloride to the solution.
- Add 2.464 g of Magnesium sulfate heptahydrate to the solution.
- Add 11.915 g of HEPES(N-2-Hydroxyethylpiperazine-N'-2-ethanesulfonic acid) to the solution.
- Add 2.1 g of Sodium bicarbonate to the solution.
- Adjust the pH to 7.8.
- Add distilled water until the volume is 1 L.
- Steriliz by autoclaving. Prepare a separate 0.1 M stock of CaCl2, and sterilize by autoclaving. Store the stocks indefinitely at room temperature. To make 1× MBS, dilute the 10× MBS stock with type 1 ultrapure water, and add CaCl2 stock to a final concentration of 0.7 mM.