Culture medium recipe
Vol.
×
Main sol. 837 #
ml
Equipment needed: Gassing station
Hungate tubes
Autoclave
Filter
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
Solution A | 870 | ml | - | - | |
Solution B | 50 | ml | - | - | |
Solution C | 20 | ml | - | - | |
Solution D | 10 | ml | - | - | |
Solution E | 50 | ml | - | - | |
1 Sparge solution A with 80% N2 and 20% CO2 gas mixture for 30 - 45 minto make it anoxic, distribute under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Solution B is autoclaved separately under 100% N2 gas atmosphere. Solution C is prepared under 80% N2 and 20% CO2 gas atmosphere. Solutions D and E are prepared under 100% N2 gas atmosphere and sterilized by filtration. To complete the medium appropriate amounts of solutions B to E are added to the sterile solution A in the sequence as indicated. Adjust pH of complete medium to 6.9, if necessary. Prior to inoculation medium is reduced by addition of 10 - 20 mg sodium dithionite per liter (e.g. from 5% (w/v) solution, freshly prepared under N2 and filter-sterilized). | |||||
2 Note: For transfers use 5 - 10% (v/v) inoculum. |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
NaCl | 20.80 | g | 23.908 | 409.104 |
KCl | 0.77 | g | 0.885 | 11.872 |
NH4Cl | 1.00 | g | 1.149 | 21.489 |
KH2PO4 | 0.10 | g | 0.115 | 0.845 |
MgSO4 x 7 H2O | 0.20 | g | 0.23 | 0.933 |
CaCl2 x 2 H2O | 0.02 | g | 0.023 | 0.156 |
Na-acetate x 3 H2O | 1.36 | g | 1.563 | - |
Modified Wolin's mineral solution | 10.00 | ml | - | - |
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 5.0e-4 | - |
Distilled water | 860.00 | ml | - | - |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
MgCl2 x 6 H2O | 10.6 | g | 212 | 1042.78 |
CaCl2 x 2 H2O | 1.5 | g | 30 | 204.061 |
Distilled water | 50.0 | ml | - | - |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
Na2CO3 | 1 | g | 50 | 471.752 |
Distilled water | 20 | ml | - | - |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
Wolin's vitamin solution | 10 | ml | - | - |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
Na2-fumarate | 8 | g | 160 | 999.775 |
Distilled water | 50 | ml | - | - |
Solution occurs in 189
other media (first defined in Medium 141).
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
Nitrilotriacetic acid | 1.50 | g | 1.5 | 7.848 | |
MgSO4 x 7 H2O | 3.00 | g | 3 | 12.172 | |
MnSO4 x H2O | 0.50 | g | 0.5 | 2.958 | |
NaCl | 1.00 | g | 1 | 17.112 | |
FeSO4 x 7 H2O | 0.10 | g | 0.1 | 0.36 | |
CoSO4 x 7 H2O | 0.18 | g | 0.18 | 1.161 | |
CaCl2 x 2 H2O | 0.10 | g | 0.1 | 0.68 | |
ZnSO4 x 7 H2O | 0.18 | g | 0.18 | 0.626 | |
CuSO4 x 5 H2O | 0.01 | g | 0.01 | 0.04 | |
AlK(SO4)2 x 12 H2O | 0.02 | g | 0.02 | 0.042 | |
H3BO3 | 0.01 | g | 0.01 | 0.162 | |
Na2MoO4 x 2 H2O | 0.01 | g | 0.01 | 0.041 | |
NiCl2 x 6 H2O | 0.03 | g | 0.03 | 0.231 | |
Na2SeO3 x 5 H2O | 0.30 | mg | 3.0e-4 | 0.001 | |
Na2WO4 x 2 H2O | 0.40 | mg | 4.0e-4 | 0.001 | |
Distilled water | 1000.00 | ml | - | - | |
1 First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Adjust final to pH 7.0 with KOH. |
Solution occurs in 351
other media (first defined in Medium 141).
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
Biotin | 2.0 | mg | 0.002 | 0.008 |
Folic acid | 2.0 | mg | 0.002 | 0.005 |
Pyridoxine hydrochloride | 10.0 | mg | 0.01 | 0.049 |
Thiamine HCl | 5.0 | mg | 0.005 | 0.015 |
Riboflavin | 5.0 | mg | 0.005 | 0.013 |
Nicotinic acid | 5.0 | mg | 0.005 | 0.041 |
Calcium D-(+)-pantothenate | 5.0 | mg | 0.005 | - |
Vitamin B12 | 0.1 | mg | 1.0e-4 | 7.4e-5 |
p-Aminobenzoic acid | 5.0 | mg | 0.005 | 0.036 |
(DL)-alpha-Lipoic acid | 5.0 | mg | 0.005 | 0.024 |
Distilled water | 1000.0 | ml | - | - |
No comments for this medium.