171 .
THERMOANAEROBACTER KIVUI MEDIUM
What is this page?
Equipment needed:
Autoclave
Gassing station
Hungate tubes
Final pH:
6.5
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
K2HPO4 | 0.22 | g | ||
|
|
KH2PO4 | 0.22 | g | ||
|
|
NaH2PO4 x H2O | 4.50 | g | ||
|
|
Na2HPO4 x 12 H2O | 6.10 | g | ||
|
|
NH4Cl | 0.31 | g | ||
|
|
(NH4)2SO4 | 0.22 | g | ||
|
|
NaCl | 0.45 | g | ||
|
|
MgSO4 x 7 H2O | 0.09 | g | ||
|
|
CaCl2 x 2 H2O
(0.1% w/v) |
6.00 | ml | ||
|
|
FeSO4 x 7 H2O solution (0.1% w/v) | 2.00 | ml | ||
|
|
Modified Wolin's mineral solution | 10.00 | ml | ||
|
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | ||
|
|
L-Cysteine HCl x H2O | 0.50 | g | ||
|
|
Na2S x 9 H2O | 0.50 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Dissolve ingredients except cysteine and sulfide, sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic, then distribute under same gas atmosphere into Hungate-type tubes or serum vials to 30% of their volume and autoclave. Add cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere. Adjust pH of complete medium to 6.5. | |||||
| 2 After inoculation pressurize cultivation vessels to one bar overpressure with sterile 80% H2 and 20% CO2 gas mixture. | |||||
Modified Wolin's mineral solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Nitrilotriacetic acid | 1.50 | g | ||
|
|
MgSO4 x 7 H2O | 3.00 | g | ||
|
|
MnSO4 x H2O | 0.50 | g | ||
|
|
NaCl | 1.00 | g | ||
|
|
FeSO4 x 7 H2O | 0.10 | g | ||
|
|
CoSO4 x 7 H2O | 0.18 | g | ||
|
|
CaCl2 x 2 H2O | 0.10 | g | ||
|
|
ZnSO4 x 7 H2O | 0.18 | g | ||
|
|
CuSO4 x 5 H2O | 0.01 | g | ||
|
|
AlK(SO4)2 x 12 H2O | 0.02 | g | ||
|
|
H3BO3 | 0.01 | g | ||
|
|
Na2MoO4 x 2 H2O | 0.01 | g | ||
|
|
NiCl2 x 6 H2O | 0.03 | g | ||
|
|
Na2SeO3 x 5 H2O | 0.30 | mg | ||
|
|
Na2WO4 x 2 H2O | 0.40 | mg | ||
|
|
Distilled water | 1000.00 | 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. | |||||
FeSO4 x 7 H2O solution (0.1% w/v) #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
FeSO4 x 7 H2O | 1 | g | ||
|
|
H2SO4
(0.1 N) |
1000 | ml | ||
| 1 The ferrous sulfate solution is not stable and should be freshly prepared. | |||||