DSMZ 700 . ALKALIPHILIC SPIROCHAETE MEDIUM

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Equipment needed:

Final pH: 9.7

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ml
Compound Amount Unit
Na2CO3 10.0 g
NaHCO3 15.0 g
NaCl 10.0 g
K2HPO4 0.2 g
NH4Cl 1.0 g
KCl 0.2 g
Na2S x 9 H2O 1.0 g
Yeast extract 0.5 g
Sucrose 5.0 g
Wolin's vitamin solution 10.0 ml
Modified Wolin's mineral solution II 1.0 ml
Distilled water 1000.0 ml
1 Prepare the medium under N2, omitting Na2CO3, Na HCO3, Na2S, sucrose and vitamin solution.
2 Boil the medium and cool under nitrogen, add the Na2CO3 and NaHCO3 and adjust the pH to 9.7 with 6N NaOH (about 15 ml). Sterilize by autoclaving and add the Na2S (neutralized), vitamins and sucrose from sterile stock solutions.

Modified Wolin's mineral solution II #

1000
Compound Amount Unit
Nitrilotriacetic acid 1.500 g
MgSO4 x 7 H2O 3.000 g
MnSO4 x H2O 0.500 g
NaCl 1.000 g
FeSO4 x 7 H2O 0.100 g
CoSO4 x 7 H2O 0.180 g
CaCl2 x 2 H2O 0.100 g
ZnSO4 x 7 H2O 0.180 g
CuSO4 x 5 H2O 0.010 g
AlK(SO4)2 x 12 H2O 0.020 g
H3BO3 0.010 g
Na2MoO4 x 2 H2O 0.010 g
NiCl2 x 6 H2O 0.025 g
Na2SeO3 x 5 H2O 0.300 mg
Distilled water 1000.000 ml
1 First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Adjust final to pH 7.0 with KOH.