DSMZ 1526e . LIMIHALOGLOBUS MEDIUM

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Equipment needed: Autoclave Filter Gassing station Hungate tubes

Final gas composition: 80 % N2 20 % CO2 read more

Final pH: 7.3 - 7.5

Select a strain (optional):

ml
Compound Amount Unit
NaCl 60.0 g
MgCl2 x 6 H2O 6.0 g
KCl 1.5 g
Na2SO4 1.0 g
NH4Cl 1.0 g
CaCl2 x 2 H2O 0.4 g
K2HPO4 0.4 g
Modified Wolin's mineral solution 10.0 ml
Sodium resazurin
(0.1% w/v)
0.5 ml
Sulfur
(powder)
10.0 g
Na2CO3 1.5 g
D-Glucose 1.0 g
Yeast extract 1.0 g
Wolin's vitamin solution (10x) 1.0 ml
L-Cysteine HCl x H2O 0.3 g
DL-Dithiothreitol
(DTT)
0.3 g
Distilled water 1000.0 ml
1 Dissolve ingredients except sulfur, carbonate, glucose, yeast extract, vitamins, cysteine and DTT, then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials containing already the appropriate amount of sulfur and autoclave at 110°C for 20 min. Add glucose, yeast extract, vitamins, cysteine and DTT from sterile anoxic stock solutions prepared under 100% N2 gas (vitamins and DTT are sterilized by filtration) and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Prior to use adjust pH of complete medium to 7.3 - 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.