663 .
DESULFITOBACTERIUM MEDIUM
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Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Final gas composition:
100 % N2
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Final pH:
7.5
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
KH2PO4 | 5.44 | g | ||
|
|
MgCl2 x 2 H2O | 0.18 | g | ||
|
|
CaCl2 x 2 H2O | 0.03 | g | ||
|
|
NH4Cl | 0.50 | g | ||
|
|
Trace element solution | 5.00 | ml | ||
|
|
Yeast extract | 1.00 | g | ||
|
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | ||
|
|
Na-pyruvate | 2.50 | g | ||
|
|
Na2S2O3 x 5 H2O | 2.00 | g | ||
|
|
Wolin's vitamin solution (10x) | 2.00 | ml | ||
|
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L-Cysteine HCl x H2O | 0.40 | g | ||
|
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Na2S x 9 H2O | 0.40 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Dissolve ingredients (except pyruvate, thiosulfate, vitamins, cysteine and sulfide), then sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add pyruvate, thiosulfate, vitamins, cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Stock solutions of pyruvate and vitamins are sterilized by filtration. Adjust medium pH to 7.5 by adding sterile anoxic 1 N NaOH. | |||||
Trace element solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Nitrilotriacetic acid
(NTA) |
12.80 | g | ||
|
|
FeCl2 x 4 H2O | 0.20 | g | ||
|
|
MnCl2 x 4 H2O | 0.10 | g | ||
|
|
CoCl2 x 6 H2O | 0.17 | g | ||
|
|
CaCl2 x 2 H2O | 0.10 | g | ||
|
|
ZnCl2 | 0.10 | g | ||
|
|
CuCl2 | 0.02 | g | ||
|
|
H3BO3 | 0.01 | g | ||
|
|
Na2MoO4 x 2 H2O | 0.01 | g | ||
|
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NiCl2 x 6 H2O | 0.03 | g | ||
|
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NaCl | 1.00 | g | ||
|
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Na2SeO3 x 5 H2O | 0.03 | g | ||
|
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Distilled water | 1000.00 | ml | ||
| 1 First dissolve NTA in 200 ml of distilled water and adjust pH to 6.5 with KOH, then dissolve mineral salts. Finally adjust pH to 6.5 with KOH and make up to 1000.00 ml. | |||||
Wolin's vitamin solution (10x) #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 20 | mg |
|
|
Folic acid | 20 | mg |
|
|
Pyridoxine hydrochloride | 100 | mg |
|
|
Thiamine HCl | 50 | mg |
|
|
Riboflavin | 50 | mg |
|
|
Nicotinic acid | 50 | mg |
|
|
Calcium D-(+)-pantothenate | 50 | mg |
|
|
Vitamin B12 | 1 | mg |
|
|
p-Aminobenzoic acid | 50 | mg |
|
|
(DL)-alpha-Lipoic acid | 50 | mg |
|
|
Distilled water | 1000 | ml |