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Modified Barth's Saline 10× Preparation and Recipe

To prepare L  Modified Barth's Saline 10×

Table 1. Recipe

NameAmountConcentrationFormula/M.W.CAS #
Sodium chloride46.752 g0.8 MNaCl/58.447647-14-5
Potassium Chloride0.745 g0.01 MKCl/74.557447-40-7
2.464 g0.01 MMgSO4·7H2O/246.4710034-99-8
HEPES(N-2-Hydroxyethylpiperazine-N'-2-ethanesulfonic acid)11.915 g0.05 MC8H18N2O4S/238.37365-45-9
Sodium bicarbonate2.1 g0.025 MNaHCO3/84.01144-55-8
  1. Prepare 0.8 L of distilled water in a suitable container.
  2. Add 46.752 g of Sodium chloride to the solution.
  3. Add 0.745 g of Potassium Chloride to the solution.
  4. Add 2.464 g of Magnesium sulfate heptahydrate to the solution.
  5. Add 11.915 g of HEPES(N-2-Hydroxyethylpiperazine-N'-2-ethanesulfonic acid) to the solution.
  6. Add 2.1 g of Sodium bicarbonate to the solution.
  7. Adjust the pH to 7.8.
  8. Add distilled water until the volume is 1 L.
  9. 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.