916 .
DESULFOFABA HANSENII MEDIUM
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Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Final pH:
7.0 - 7.2
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Solution A | 912 | ml | ||
|
|
Solution B | 50 | ml | ||
|
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Solution C | 25 | ml | ||
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Solution D | 10 | ml | ||
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Solution E | 1 | ml | ||
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Solution F | 10 | ml | ||
| 1 Sparge solution A with 80% N2 and 20% CO2 gas mixture for 30 – 45 min to make it anoxic, then dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Solutions B and D are autoclaved under 100% N2 gas atmosphere. Solution C is autoclaved separately under 80% N2 and 20% CO2 gas atmosphere. Solutions E and F are prepared under 100% N2 gas and sterilized by filtration. To complete the medium appropriate amounts of solutions B to F are added to the sterile solution A in the sequence as indicated. The pH of the complete medium should be 7.0 - 7.2. | |||||
| 2 Note: Addition of 10 - 20 mg sodium dithionite per liter (e.g. from 5% (w/v) solution, freshly prepared under N2 and filter-sterilized) may stimulate growth at the beginning. | |||||
Solution A #
912
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
NaCl | 20.00 | g |
|
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KCl | 0.67 | g |
|
|
NH4Cl | 0.10 | g |
|
|
KH2PO4 | 0.01 | g |
|
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MgSO4 x 7 H2O | 0.02 | g |
|
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Na2SO4 | 1.42 | g |
|
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Trace element solution SL-10 | 1.00 | ml |
|
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Selenite-tungstate solution | 1.00 | ml |
|
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Sodium resazurin
(0.1% w/v) |
0.50 | ml |
|
|
Distilled water | 910.00 | ml |
Solution B #
50
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
MgCl2 x 6 H2O | 10.60 | g |
|
|
CaCl2 x 2 H2O | 1.52 | g |
|
|
Distilled water | 50.00 | ml |
Solution F #
10
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
FeCl2 x 4 H2O | 0.52 | g | ||
|
|
Distilled water | 10.00 | ml | ||
| 1 Adjust to pH 2 with HCl. | |||||
Trace element solution SL-10 #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
HCl
(25%) |
10.0 | ml | ||
|
|
FeCl2 x 4 H2O | 1.5 | g | ||
|
|
ZnCl2 | 70.0 | mg | ||
|
|
MnCl2 x 4 H2O | 100.0 | mg | ||
|
|
H3BO3 | 6.0 | mg | ||
|
|
CoCl2 x 6 H2O | 190.0 | mg | ||
|
|
CuCl2 x 2 H2O | 2.0 | mg | ||
|
|
NiCl2 x 6 H2O | 24.0 | mg | ||
|
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Na2MoO4 x 2 H2O | 36.0 | mg | ||
|
|
Distilled water | 990.0 | ml | ||
| 1 First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml. | |||||
Selenite-tungstate solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
NaOH | 0.5 | g |
|
|
Na2SeO3 x 5 H2O | 3.0 | mg |
|
|
Na2WO4 x 2 H2O | 4.0 | mg |
|
|
Distilled water | 1000.0 | ml |
Wolin's vitamin solution (10x) #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 20 | mg |
|
|
Folic acid | 20 | mg |
|
|
Pyridoxine hydrochloride | 100 | mg |
|
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Thiamine HCl | 50 | mg |
|
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Riboflavin | 50 | mg |
|
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Nicotinic acid | 50 | mg |
|
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Calcium D-(+)-pantothenate | 50 | mg |
|
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Vitamin B12 | 1 | mg |
|
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p-Aminobenzoic acid | 50 | mg |
|
|
(DL)-alpha-Lipoic acid | 50 | mg |
|
|
Distilled water | 1000 | ml |