Solution A
Nitrilotriacetic acid 1.5 g, MgSO4 * 7H2O 3.0 g, MnSO4 * 2 H2O 0.5 g, NaCl
1.0 g, FeSO4 * 7 H2O 0.1 g, CaSO4 * 7 H2O 0.18g, CaCl2 * 2H2O 0.1 g,
ZnSO4 * 7H2O 0.18 g, CuSO4 * 5H2O 0.01 g, KAl(SO4)2 * 12H2O 0.02 g,
H3BO3 0.01 g, Na2MoO4 * 2 H2O 0.01 g, NiCl2 * 6 H2O 0.025 g, Na2SeO3 0.3
mg, Distilled water 1.0 L.
Nitrilotriacetic acid 1.5 g, MgSO4 * 7H2O 3.0 g, MnSO4 * 2 H2O 0.5 g, NaCl
1.0 g, FeSO4 * 7 H2O 0.1 g, CaSO4 * 7 H2O 0.18g, CaCl2 * 2H2O 0.1 g,
ZnSO4 * 7H2O 0.18 g, CuSO4 * 5H2O 0.01 g, KAl(SO4)2 * 12H2O 0.02 g,
H3BO3 0.01 g, Na2MoO4 * 2 H2O 0.01 g, NiCl2 * 6 H2O 0.025 g, Na2SeO3 0.3
mg, Distilled water 1.0 L.
COMPOSITION
NH4Cl 1.0 g, KH2PO4 0.33 g, K2HPO4 0.45 g, MgSO4 * 7H2O 0.1 g, Solution A 20 ml, Solution B 20 ml, Yeast extract
2.00 g, Fructose 1.0 g, Resazurin 1.0 mg NaHCO3 10.0 g, Cysteine hydrochloride 0.5 g, Na2S * 9H2O 0.5 g, Distilled
water 1.0 L.
NH4Cl 1.0 g, KH2PO4 0.33 g, K2HPO4 0.45 g, MgSO4 * 7H2O 0.1 g, Solution A 20 ml, Solution B 20 ml, Yeast extract
2.00 g, Fructose 1.0 g, Resazurin 1.0 mg NaHCO3 10.0 g, Cysteine hydrochloride 0.5 g, Na2S * 9H2O 0.5 g, Distilled
water 1.0 L.
| Acetobacterium medium |
Solution B
Biotin 2.0 mg, Folic acid 20 mg, Pyridoxine-HCl 10 mg, Thiamine-HCl 5.0
mg, Riboflavin 5.0 mg, Nicotinic acid 5.0 mg, DL-Calcium panthotenate 5.0
mg, Vitamin B12 0.1 mg, p-Aminobenzoic acid 5.0 mg, Lipoic acid 5.0 mg,
Distilled water 1.0 L
Biotin 2.0 mg, Folic acid 20 mg, Pyridoxine-HCl 10 mg, Thiamine-HCl 5.0
mg, Riboflavin 5.0 mg, Nicotinic acid 5.0 mg, DL-Calcium panthotenate 5.0
mg, Vitamin B12 0.1 mg, p-Aminobenzoic acid 5.0 mg, Lipoic acid 5.0 mg,
Distilled water 1.0 L
PROCEDURE
First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Final ph 7.0
Dissolve ingredients except NaHCO3, fructose, cysteine and sodium sulphide, bring to the boil for a few minutes and
cool to room temperature under N2 + CO2 (80 + 20) gas mixture. Add NaHCO3 and equilibrate medium with the gas
until a pH of approximately 7.4 is reached. Then distribute and autoclave under the same gas.
Before use adjust the pH to 8.2 by adding sterile anaerobic Na2CO3 solution (0.25 ml of 5% Na2CO3 per 10 ml
medium) and add fructose, cysteine & sodium sulphide from anaerobic sterile stock solutions.
First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Final ph 7.0
Dissolve ingredients except NaHCO3, fructose, cysteine and sodium sulphide, bring to the boil for a few minutes and
cool to room temperature under N2 + CO2 (80 + 20) gas mixture. Add NaHCO3 and equilibrate medium with the gas
until a pH of approximately 7.4 is reached. Then distribute and autoclave under the same gas.
Before use adjust the pH to 8.2 by adding sterile anaerobic Na2CO3 solution (0.25 ml of 5% Na2CO3 per 10 ml
medium) and add fructose, cysteine & sodium sulphide from anaerobic sterile stock solutions.
(c) Costin Stoica
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