1095 .
NATRANAEROBIUS MEDIUM
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* Medium for Natranaerobius sp. DSM 23380
Adjust pH of medium to 9.5 and replace sucrose with 5.0 g/l D-glucose added from a sterile anoxic stock solution after autoclaving.
Growth conditions: anaerobic
45 °C
Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Final gas composition:
100 % N2
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Final pH:
* 9.5
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaCl | 100.0 | g | ||
|
|
KH2PO4 | 0.2 | g | ||
|
|
MgCl2 | 0.1 | g | ||
|
|
NH4Cl | 0.5 | g | ||
|
|
Yeast extract
(OXOID) |
10.0 | g | ||
|
|
Tryptone
(BD Bacto) |
10.0 | g | ||
|
|
Trace element solution SL-10 | 1.0 | ml | ||
|
|
Na2CO3 | 68.0 | g | ||
|
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NaHCO3 | 38.0 | g | ||
|
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L-Cysteine HCl x H2O | 0.7 | g | ||
|
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Wolin's vitamin solution (10x) | 1.0 | ml | ||
| Sucrose* | 5.0 | g | |||
|
|
D-Glucose | 5.0 | g | ||
|
|
Distilled water | 1000.0 | ml | ||
| 1 Dissolve ingredients (except carbonates, cysteine, sucrose and vitamins), then sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Add carbonate, hydrogencarbonate and cysteine and adjust pH to 8.5. Dispense medium under 100% N2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After autoclaving add sucrose and vitamins from sterile anoxic stock solutions prepared under 100% N2 gas and sterilized by filtration. | |||||
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. | |||||
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 |