1028 .
THERMINCOLA MEDIUM (CO)
What is this page?
Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Serum bottle
Final pH:
8.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NH4Cl | 1.00 | g | ||
|
|
MgCl2 x 6 H2O | 0.33 | g | ||
|
|
CaCl2 x 2 H2O | 0.10 | g | ||
|
|
KCl | 0.33 | g | ||
|
|
KH2PO4 | 0.50 | g | ||
|
|
Wolfe's mineral elixir | 1.00 | ml | ||
|
|
Na-acetate | 0.20 | g | ||
|
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | ||
|
|
NaHCO3 | 0.50 | g | ||
|
|
Na2CO3 | 0.50 | g | ||
|
|
Na2S x 9 H2O | 1.00 | g | ||
|
|
Yeast extract | 0.20 | g | ||
|
|
Wolin's vitamin solution (10x) | 2.00 | ml | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Dissolve ingredients except carbonates, sulfide, yeast extract and vitamins, then sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Add carbonates and sulfide to the medium, dispense under 100% carbon monoxide gas atmosphere into anoxic Hungate-type tubes or serum vials (e.g., 10 ml medium in 50 ml bottles) and autoclave. Prior to inoculation add yeast extract and vitamins (sterilized by filtration) from sterile anoxic stock solutions prepared under 100% N2 gas. If the medium is not completely reduced after autoclaving (remains pink) add 0.30 g/l sodium sulfide from a sterile anoxic stock solution prepared under 100% N2 gas atmosphere. Adjust pH of complete medium to 8.0 with a sterile anoxic solution of 1 N HCl. | |||||
Wolfe's mineral elixir #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
MgSO4 x 7 H2O | 30.00 | g | ||
|
|
MnSO4 x H2O | 5.00 | g | ||
|
|
NaCl | 10.00 | g | ||
|
|
FeSO4 x 7 H2O | 1.00 | g | ||
|
|
CoCl2 x 6 H2O | 1.80 | g | ||
|
|
CaCl2 x 2 H2O | 1.00 | g | ||
|
|
ZnSO4 x 7 H2O | 1.80 | g | ||
|
|
CuSO4 x 5 H2O | 0.10 | g | ||
|
|
AlK(SO4)2 x 12 H2O | 0.18 | g | ||
|
|
H3BO3 | 0.10 | g | ||
|
|
Na2MoO4 x 2 H2O | 0.10 | g | ||
|
|
(NH4)2Ni(SO4)2 x 6 H2O | 2.80 | g | ||
|
|
Na2WO4 x 2 H2O | 0.10 | g | ||
|
|
Na2SeO4 | 0.10 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 First adjust pH to 1.0 with diluted H2SO4, then add and dissolve the salts. | |||||
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 |