DSMZ 479a . METHANOHALOPHILUS PROFUNDI MEDIUM

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Equipment needed: Autoclave Gassing station Hungate tubes

Final gas composition: 100 % N2 read more

Final pH: 7.5

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ml
Compound Amount Unit
NaCl 120.00 g
KCl 1.50 g
MgCl2 x 6 H2O 6.00 g
CaCl2 x 2 H2O 0.40 g
NH4Cl 1.00 g
K2HPO4 x 3 H2O 0.40 g
Modified Wolin's mineral solution 10.00 ml
Yeast extract
(OXOID)
2.00 g
Trypticase peptone
(BD BBL)
2.00 g
HEPES
(SIGMA H3375)
5.00 g
Sodium resazurin
(0.1% w/v)
0.50 ml
Trimethylamine-HCl 2.00 g
2-Mercaptoethanesulfonic acid
(coenzyme M)
0.20 g
Na2S x 9 H2O 0.25 g
Distilled water 1000.00 ml
1 Dissolve ingredients except trimethylamine, coenzyme M and sulfide. Adjust pH to 7.5 and sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic; then dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add trimethylamine, coenzyme M and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Adjust pH of complete medium to 7.5, if necessary.

Modified Wolin's mineral solution #

1000
Compound Amount Unit
Nitrilotriacetic acid 1.50 g
MgSO4 x 7 H2O 3.00 g
MnSO4 x H2O 0.50 g
NaCl 1.00 g
FeSO4 x 7 H2O 0.10 g
CoSO4 x 7 H2O 0.18 g
CaCl2 x 2 H2O 0.10 g
ZnSO4 x 7 H2O 0.18 g
CuSO4 x 5 H2O 0.01 g
AlK(SO4)2 x 12 H2O 0.02 g
H3BO3 0.01 g
Na2MoO4 x 2 H2O 0.01 g
NiCl2 x 6 H2O 0.03 g
Na2SeO3 x 5 H2O 0.30 mg
Na2WO4 x 2 H2O 0.40 mg
Distilled water 1000.00 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.