Supplement medium with 0.80 g/l yeast extract and replace Na-acetate with 1.10 g/l Na-L-lactate.
Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
NaCl | 1.20 | g | ||
|
KCl | 0.30 | g | ||
|
NH4Cl | 0.30 | g | ||
|
KH2PO4 | 0.20 | g | ||
|
Na2SO4 | 0.30 | g | ||
|
MgCl2 x 6 H2O | 0.40 | g | ||
|
CaCl2 x 2 H2O | 0.15 | g | ||
|
Trace element solution SL-11 | 1.00 | ml | ||
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | ||
|
NaHCO3 | 0.60 | g | ||
|
KNO3 | 0.50 | g | ||
|
Wolin's vitamin solution (10x) | 1.00 | ml | ||
|
Seven vitamins solution | 1.00 | ml | ||
Na-acetate* | 0.50 | g | |||
|
Yeast extract | 0.80 | g | ||
|
Na-(L)-lactate | 1.10 | g | ||
|
Distilled water | 1000.00 | ml | ||
1 Dissolve ingredients except bicarbonate, nitrate, vitamins and acetate. Sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic, adjust pH to 7.0 - 7.4, then dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add nitrate, vitamins and acetate from sterile anoxic stock solutions prepared under 100% N2 gas and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas atmosphere. Vitamins should be sterilized by filtration. The pH of the complete medium should be adjusted to 7.4 - 7.8, if necessary. |
Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
Na2-EDTA x 2 H2O | 5.2 | g | ||
|
FeCl2 x 4 H2O | 1.5 | g | ||
|
ZnCl2 | 70.0 | mg | ||
|
MnCl2 x 4 H2O | 100.0 | mg | ||
|
H3BO3 | 6.0 | mg | ||
|
CoCl2 x 6 H2O | 190.0 | mg | ||
|
CuCl2 x 2 H2O | 2.0 | mg | ||
|
NiCl2 x 6 H2O | 24.0 | mg | ||
|
Na2MoO4 x 2 H2O | 36.0 | mg | ||
|
Distilled water | 1000.0 | ml | ||
1 Dissolve EDTA in 800 ml distilled water, adjust pH to 7 using 2 N NaOH and add ferrous chloride. After ferrous chloride has dissolved add other compounds. Finally adjust pH to 6.0 and bring volume to 1000 ml. |
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 |
Compound | Amount | Unit | |
---|---|---|---|
|
Vitamin B12 | 100 | mg |
|
p-Aminobenzoic acid | 80 | mg |
|
D-(+)-biotin | 20 | mg |
|
Nicotinic acid | 200 | mg |
|
Calcium pantothenate | 100 | mg |
|
Pyridoxine hydrochloride | 300 | mg |
|
Thiamine-HCl x 2 H2O | 200 | mg |
|
Distilled water | 1000 | ml |