NH4Cl……………………………….....…................... 1.0 g
KH
2PO4 ………………………………...................… 0.33 g
K
2HPO4 ………...….….......................................… 0.45 g
MgSO4 * 7H
2O ……….............................….……… 0.1 g
Solution A ………………………….......………...….. 20 ml
Solution B ………………………….......………...….. 20 ml
Yeast extract ……………………………….........…. 2.00 g
Fructose ……………………………………...……..... 1.0 g
Resazurin ............................................................1.0 mg
NaHCO
3 ............................................................... 10.0 g
Cysteine hydrochloride .......................................... 0.5 g
Na
2S * 9H2O ........................................................... 0.5 g
Distilled water......................................................... 1.0 L


Solution A
Nitrilotriacetic acid ................................................. 1.5 g
MgSO
4 * 7H2O …………………............................... 3.0 g
MnSO
4 * 2 H2O ....................................................... 0.5 g
NaCl ...................................................................... 1.0 g
FeSO
4 * 7 H2O ....................................................... 0.1 g
CaSO
4 * 7 H2O ...................................................... 0.18g
CaCl
2 * 2H2O .......................................................... 0.1 g
ZnSO
4 * 7H2O ....................................................... 0.18 g
CuSO
4 * 5H2O ...................................................... 0.01 g
KAl(SO
4)2 * 12H2O ................................................ 0.02 g
H
3BO3 ................................................................... 0.01 g
Na
2MoO4 * 2 H2O .................................................. 0.01 g
NiCl
2 * 6 H2O ...................................................... 0.025 g
Na
2SeO3 .............................................................. 0.3 mg
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
First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Final ph 7.0

Dissolve ingredients except NaHCO
3, fructose, cysteine and sodium sulphide, bring to the boil for a few minutes and
cool to room temperature under N
2 + 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 Na
2CO3 solution (0.25 ml of 5% Na2CO3 per 10 ml
medium) and add fructose, cysteine & sodium sulphide from anaerobic sterile stock solutions.
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