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Main sol. 3

Internal Solution-ID: S5

Compound Amount Unit Conc. [g/L] Conc. [mM]
Glucose 5.00 g 5 27.753
Mannitol 5.00 g 5 27.447
CaCl2 x 2 H2O 0.10 g 0.1 0.68
MgSO4 x 7 H2O 0.10 g 0.1 0.406
Na2MoO4 x 2 H2O 5.00 mg 0.005 0.021
K2HPO4 0.90 g 0.9 5.167
KH2PO4 0.10 g 0.1 0.735
FeSO4 x 7 H2O 0.01 g 0.01 0.036
CaCO3 5.00 g 5 49.955
Agar 15.00 g 15 44.599
Distilled water 950.00 ml - -
1 Adjust pH to 7.3.
2 Sterilize glucose and mannitol separately (in 50 ml H2O) and add to the medium after autoclaving. Calcium carbonate in the medium serves as a buffer. The calcium carbonate will settle in agar plates before the agar has set, producing an opaque layer in the bottom. As the strain grows and acid is produced this will react with the calcium carbonate, causing it to dissolve and form zones of clearing immediately below the colonies.
3 AZOTOBACTER MEDIUM