505 .
FORMIVIBRIO MEDIUM
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* Medium for Youngiibacter multivorans DSM 6139
Use instead of citrate 10 mM sodium cinnamate (or 0.2% sucrose or 0.2% sodium pyruvate) as substrate and supplement the medium with 0.05 g/l of yeast extract. Sterilize substrates separately. Adjust pH of the complete medium to 7.7 - 7.9.
Growth conditions: anaerobic
30 °C
Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Final pH:
* 7.7 - 7.9
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Solution A | 942 | ml | ||
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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. Solutions C and E are autoclaved under 100% N2 gas. Solution D is prepared under 100% N2 gas and sterilized by filtration. 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 7.5 - 7.8 with a sterile anoxic stock solution of Na2CO3, if necessary. | |||||
Solution A #
942
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
KH2PO4 | 1.40 | g |
|
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NH4Cl | 0.50 | g |
|
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MgCl2 x 6 H2O | 0.20 | g |
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CaCl2 x 2 H2O | 0.15 | g |
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Sodium resazurin
(0.1% w/v) |
0.50 | ml |
|
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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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Yeast extract | 0.05 | g |
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Distilled water | 940.00 | ml |
Solution C #
20
| Compound | Amount | Unit | |
|---|---|---|---|
| Trisodium citrate x 2 H2O* | 3.0 | g | |
|
|
Sodium cinnamate | 1.7 | g |
|
|
Distilled water | 20.0 | 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 | ||
|
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MnCl2 x 4 H2O | 100.0 | mg | ||
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H3BO3 | 6.0 | mg | ||
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CoCl2 x 6 H2O | 190.0 | mg | ||
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|
CuCl2 x 2 H2O | 2.0 | mg | ||
|
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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 |
|
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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 |
|
|
Nicotinic acid | 50 | mg |
|
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Calcium D-(+)-pantothenate | 50 | mg |
|
|
Vitamin B12 | 1 | mg |
|
|
p-Aminobenzoic acid | 50 | mg |
|
|
(DL)-alpha-Lipoic acid | 50 | mg |
|
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Distilled water | 1000 | ml |