503 .
ANAEROBIC FRESHWATER (FWM) MEDIUM
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* Medium for Fusibacter fontis DSM 28450
Supplement medium with 2.00 g/l yeast extract added from an anoxic stock solution sterilized by filtration.
Growth conditions: anaerobic
30 °C
Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Final pH:
7.2 - 7.4
Select a strain (optional):
Main sol. 503 #
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Solution A | 942 | ml | ||
|
|
Solution B | 30 | ml | ||
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Solution C | 20 | ml | ||
|
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Solution D | 1 | ml | ||
|
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Solution E | 10 | ml | ||
| 1 Sparge solution A with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic, distribute under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Solution B is autoclaved separately under 80% N2 and 20% CO2 gas atmosphere. Solution C and D are prepared under 100% N2 gas and sterilized by filtration. Solution E is autoclaved under 100% N2 gas. To complete the medium appropriate amounts of solutions B to E are added to the sterile solution A in the sequence as indicated. Adjust pH of complete medium to 7.2 - 7.4, if necessary. | |||||
Solution A #
942
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
KH2PO4 | 0.20 | g |
|
|
NH4Cl | 0.25 | g |
|
|
NaCl | 1.00 | g |
|
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MgCl2 x 6 H2O | 0.40 | g |
|
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KCl | 0.50 | g |
|
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CaCl2 x 2 H2O | 0.15 | 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 |
|
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Yeast extract | 2.00 | g |
|
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Distilled water | 940.00 | ml |
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 | ||
|
|
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 |
Seven vitamins solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Vitamin B12 | 100 | mg |
|
|
p-Aminobenzoic acid | 80 | mg |
|
|
D-(+)-biotin | 20 | mg |
|
|
Nicotinic acid | 200 | mg |
|
|
Calcium pantothenate | 100 | mg |
|
|
Pyridoxine hydrochloride | 300 | mg |
|
|
Thiamine-HCl x 2 H2O | 200 | mg |
|
|
Distilled water | 1000 | ml |