Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
Solution A | 952 | ml | ||
|
Solution B | 10 | ml | ||
|
Solution C | 10 | ml | ||
|
Solution D | 30 | ml | ||
|
Solution E | 1 | ml | ||
1 Solution A is sparged with 80% N2 and 20% CO2 gas mixture to reach a pH below 6 (at least 30 min), then distributed under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclaved. Solutions B and C are autoclaved separately under 100% N2 gas. Solution D is autoclaved under 80% N2 and 20% CO2 gas atmosphere. Solution E is prepared under 100% N2 gas atmosphere and sterilized by filtration. Solutions B to E are added to the sterile, cooled solution A in appropriate amounts in the sequence as indicated. Adjust pH of final medium to 7.2, if necessary. | |||||
2 For aerobic growth prepare medium under air. Omit carbonate and KNO3. 3,5-Dihydroxybenzoic acid or Na-benzoate as substrate can be replaced with 10 mM Na-malate. Adjust medium pH to 7.2. |
Compound | Amount | Unit | |
---|---|---|---|
|
Na2SO4 | 3.00 | g |
|
KH2PO4 | 0.20 | g |
|
NH4Cl | 0.30 | g |
|
NaCl | 1.00 | g |
|
MgCl2 x 6 H2O | 0.40 | g |
|
KCl | 0.50 | g |
|
CaCl2 x 2 H2O | 0.15 | g |
|
Trace element solution SL-10 | 1.00 | ml |
|
Selenite-tungstate solution | 1.00 | ml |
|
Distilled water | 950.00 | ml |
Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
HCl
(25%) |
10.0 | ml | ||
|
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 | 990.0 | ml | ||
1 First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml. |
Compound | Amount | Unit | |
---|---|---|---|
|
NaOH | 0.5 | g |
|
Na2SeO3 x 5 H2O | 3.0 | mg |
|
Na2WO4 x 2 H2O | 4.0 | mg |
|
Distilled water | 1000.0 | 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 |