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| Composition | capacity | unit |
|---|---|---|
| Thiamine | 5.0 | mg |
| MgSO4·7H2O | 0.1 | g |
| Wolfe''s vitamin solution (see below) | 10.0 | ml |
| KH2PO4 | 0.4 | g |
| Resazurin solution (0.01 %) | 1.0 | ml |
| Wolfe''s vitamin solution: Folic acid | 2.0 | mg |
| Distilled water | 955.0 | ml |
| Fructose (25 ml of a 20 % solution) | 5.0 | g |
| Yeast extract | 1.0 | g |
| NH4Cl | 1.0 | g |
| Wolfe''s mineral solution(see below) | 10.0 | ml |
| K2HPO_4 _ | 0.4 | g |
| Riboflavin | 5.0 | mg |
| NaHCO3 | 3.0 | g |
| Nicotinic acid | 5.0 | mg |
| Pantothenic acid | 5.0 | mg |
| Vitamin B_12 _ | 0.1 | mg |
| p-Aminobenzoic acid | 5.0 | mg |
| Thioctic acid | 5.0 | mg |
| Distilled water | 1.0 | L |
| Wolfe''s mineral solution: Nitriloacetic acid | 1.5 | g |
| MgSO4·7H2O | 3.0 | g |
| FeSO4·7H2O | 0.1 | g |
| Pyridoxine hydrochloride | 10.0 | mg |
| Biotin | 2.0 | mg |
| CuSO4·5H2O | 10.0 | mg |
| AlK(SO4)2·12H2O | 10.0 | mg |
| NaCl | 1.0 | g |
| ZnSO4·H2O | 0.1 | g |
| CoCl2·6H2O | 0.1 | g |
| CaCl2 | 0.1 | g |
| MnSO4·H2O | 0.5 | g |
| Na2MoO4·2H2O | 10.0 | mg |
| Distilled water | 1.0 | L |
| H3BO3 | 10.0 | mg |
** ph : -
** memo : Add nitrilotriacetic acid to approximately 500 ml of water and adjust to pH 6.5 with KOH to dissolve the compound. Bring volume to 1.0 L with remaining water and add remaining compounds one at a time. Mix all of the ingredients with the exception of the 25 ml of filter-sterilized fructose. Bring to boil to drive oxygen out. Add 40.0 ml of a solution containing 1.25 % cysteine HCl and 1.25 % Na_2_S*9H_2_O and autoclave at 121°C for 15 minutes. After autoclaving and while still hot, bubble through oxygen-free, 90 % N_2 _and 10 % CO_2_ to maintain anaerobiosis. When the medium has cooled to about 50℃, add the sterile fructose. Adjust pH if necessary to 6.7, tube aseptically under anaerobic" conditions and cap with rubber stoppers.
** additional actions :