Medium data
Search for medium no. = [221].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.
221 THERMOANAEROBACTER ETHANOLICUS MEDIUM
| KH2PO4 | 1.5 | g |
| Na2HPO4·12H2O | 4.2 | g |
| NH4Cl | 0.5 | g |
| MgCl2·6H2O | 0.18 | g |
| Yeast extract (BD-Difco) | 2.0 | g |
| Glucose | 8.0 | g |
| Vitamin solution (see below) | 0.5 | ml |
| Wolfe's modified mineral elixir (see below) | 5.0 | ml |
| Resazurin | 1.0 | mg |
| Reducing solution (see below) | 40.0 | ml |
| Distilled water | 960.0 | ml |
Mix ingredients, except reducing solution, and bring to the boil while gassing with N2-H2 (95:5) gas mixture. When the color turns from blue to reddish pink, add the reducing solution during continuous gassing and boiling. Anaerobically dispense the medium in tubes under a N2-H2 (95:5) atmosphere. Stopper the tubes and autoclave.
Vitamin solution:
| Biotin | 20.0 | mg |
| p-Aminobenzoic acid | 50.0 | mg |
| Folic acid | 20.0 | mg |
| Calcium pantothenate | 50.0 | mg |
| Nicotinic acid | 50.0 | mg |
| Vitamin B12 | 1.0 | mg |
| Thiamine·HCl | 5.0 | mg |
| Pyridoxine·HCl | 100.0 | mg |
| Thioctic acid | 50.0 | mg |
| Riboflavin | 5.0 | mg |
| Distilled water | 500.0 | ml |
Store at -20C or 4C in the absence of light.
Wolfe's modified mineral elixir:
| Nitrilotriacetic acid | 1.5 | g |
| MgSO4·7H2O | 3.0 | g |
| MnSO4·xH2O | 0.5 | g |
| NaCl | 1.0 | g |
| FeSO4·7H2O | 0.1 | g |
| Co(NO3)2·6H2O | 0.1 | g |
| CaCl2 | 0.1 | g |
| ZnSO4·7H2O | 0.1 | g |
| CuSO4·5H2O | 0.01 | g |
| AlK(SO4)2 | 0.01 | g |
| H3BO3 | 0.01 | g |
| Na2MoO4·2H2O | 0.01 | g |
| Na2SeO3 | 1.0 | mg |
| Distilled water | 1.0 | L |
Dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH. Then proceed to add minerals in the order listed.
Reducing solution:
| 0.2 N NaOH | 200.0 | ml |
| Na2S·9H2O | 2.5 | g |
| L-Cysteine·HCl·H2O | 2.5 | g |
Boil NaOH solution and cool down under a N2-H2 (95:5) atmosphere, then add Na2S·9H2O and L-cysteine·HCl·H2O to the solution. Autoclave it under a N2-H2 (95:5) atmosphere.
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