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Main sol. J462

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Internal Solution-ID: S4209

Equipment needed: Hungate tubes Filter Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 20.00 g 3333.33 57038.6
Na2SO4 4.00 g 666.667 4693.51
MgCl2 x 6 H2O 3.00 g 500 2459.38
CaCl2 x 2 H2O 0.15 g 25 170.051
NH4Cl 0.75 g 125 2336.89
KH2PO4 0.60 g 100 734.829
KBr 0.09 g 15 126.05
KCl 0.50 g 83.333 1117.82
NaHCO3 2.50 g 416.667 4959.9
Trace element solution 1.00 ml - -
Vitamin solution 1.00 ml - -
Thiamine solution 1.00 ml - -
Vitamin B12 solution 0.05 g/L 1.00 ml - -
Riboflavin solution 0.025 g/L 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
Sodium lactate 2.25 g 375 3346.42
Na2S x 9 H2O 0.30 g 50 208.175
Resazurin 1.00 mg 0.167 0.727
1 Dissolve components except NaHCO3, Vitamin solution, Thiamine solution, Vitamin B12 solution, sodium lactate and Na2S x 9H2O in 900 ml distilled water and adjust pH to 7.0. Bring to a boil and cool down under a N2-CO2 (4:1, v/v) gas stream. Distribute the medium into culture vessels (e.g., 9 ml of the medium in Hungate tubes) under a N2-CO2 (4:1, v/v) gas mixture, seal with butyl rubber stoppers and autoclave. Separately autoclave 0.2 M sodium lactate under a N2 atmosphere. After cooling, add 1/9 volume of the sodium lactate solution, and filter-sterilized 8% NaHCO3 solution, Vitamin solution, Thiamine solution and Vitamin B12 solution to the medium. Readjust pH to 7.0, if necessary. Prior to use, reduce the medium with Na2S x 9H2O (3% solution, autoclaved and stocked under N2).
J462 SALTWATER MEDIUM WITH LACTATE