534 .
MODIFIED SME MEDIUM
What is this page?
Equipment needed:
Autoclave
Filter
Final pH:
6.6
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaCl | 30.00 | g | ||
|
|
MgSO4 x 7 H2O | 7.00 | g | ||
|
|
MgCl2 x 6 H2O | 5.50 | g | ||
|
|
KCl | 0.65 | g | ||
|
|
NaBr | 0.10 | g | ||
|
|
NH4Cl | 0.15 | g | ||
|
|
K2HPO4 | 0.15 | g | ||
|
|
CaCl2 x 2 H2O | 0.50 | g | ||
|
|
Trace element solution | 10.00 | ml | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Sparge medium with 100% N2 gas mixture for 20 min to make it anoxic. | |||||
| 2 Adjust pH to 6.5-6.8 with 1N H2SO4. | |||||
| 3 Fill 10 ml of medium in serum bottles (100 ml) under a protective H2/CO2 atmosphere and tightly stopper. | |||||
| 4 Autoclave at 121 °C for 15 min. | |||||
| 5 after autoclaving add the following filter-sterilized solutions. | |||||
|
|
Solution A
, 0.3 ml/10ml
(gassed with CO2) |
30.00 | ml/l | ||
|
|
Solution B
, 0.3 ml/10ml
(gassed with N2) |
30.00 | ml/l | ||
| 6 apply xx bar H2/CO2 (80%/20%) + yy % O2 (Note: strain specific conditions need to be applied) | |||||
| 7 For more details on the medium see the original description (Huber et al. System. Appl. Microbiol. 15:340-351 (1992)). | |||||
Trace element solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Modified Wolin's mineral solution II | 1000 | ml | ||
| 1 To 1000.0 ml trace element solution in medium 141 add: | |||||
|
|
(NH4)2Ni(SO4)2 | 2 | g | ||
|
|
Na2WO4 | 10 | mg | ||
|
|
Na2SeO4 | 10 | mg | ||
Solution A #
30
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaHCO3 | 2 | g | ||
|
|
Distilled water | 30 | ml | ||
| 1 prepared under CO2 in an ice-bath, filter-sterilise | |||||
Solution B #
30
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Na2S2O3 x 5 H2O | 2 | g | ||
|
|
Distilled water | 30 | ml | ||
| 1 Prepared under N2,autoclave 121°C, 15 min | |||||
Modified Wolin's mineral solution II #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Nitrilotriacetic acid | 1.500 | g | ||
|
|
MgSO4 x 7 H2O | 3.000 | g | ||
|
|
MnSO4 x H2O | 0.500 | g | ||
|
|
NaCl | 1.000 | g | ||
|
|
FeSO4 x 7 H2O | 0.100 | g | ||
|
|
CoSO4 x 7 H2O | 0.180 | g | ||
|
|
CaCl2 x 2 H2O | 0.100 | g | ||
|
|
ZnSO4 x 7 H2O | 0.180 | g | ||
|
|
CuSO4 x 5 H2O | 0.010 | g | ||
|
|
AlK(SO4)2 x 12 H2O | 0.020 | g | ||
|
|
H3BO3 | 0.010 | g | ||
|
|
Na2MoO4 x 2 H2O | 0.010 | g | ||
|
|
NiCl2 x 6 H2O | 0.025 | g | ||
|
|
Na2SeO3 x 5 H2O | 0.300 | mg | ||
|
|
Distilled water | 1000.000 | ml | ||
| 1 First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Adjust final to pH 7.0 with KOH. | |||||