Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
NH4Cl | 1.00 | g | ||
|
NaCl | 0.10 | g | ||
|
MgCl2 x 6 H2O | 0.10 | g | ||
|
CaCl2 x 2 H2O | 0.05 | g | ||
|
K2HPO4 x 3 H2O | 0.40 | g | ||
|
Modified Wolin's mineral solution | 10.00 | ml | ||
|
Yeast extract | 0.75 | g | ||
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | ||
|
Na2CO3 | 1.00 | g | ||
|
Cellobiose | 4.00 | g | ||
|
Cellulose
, Avicel SIGMA
optional |
2.00 | g | ||
|
Wolin's vitamin solution (10x) | 1.00 | ml | ||
|
Na2S x 9 H2O | 0.25 | g | ||
|
Distilled water | 1000.00 | ml | ||
1 Dissolve ingredients (except carbonate, cellobiose, vitamins and sulfide), then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After sterilization add cellobiose, vitamins and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Stock solutions of cellobiose and vitamins should be sterilized by filtration. Adjust pH of complete medium to 7.0, if necessary. | |||||
2 Note: Some strains can be adapted to cellulose as substrate using 2.00 g/l Avicel microcrystalline cellulose (SIGMA). |
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 | |||
---|---|---|---|---|---|
|
Nitrilotriacetic acid | 1.50 | g | ||
|
MgSO4 x 7 H2O | 3.00 | g | ||
|
MnSO4 x H2O | 0.50 | g | ||
|
NaCl | 1.00 | g | ||
|
FeSO4 x 7 H2O | 0.10 | g | ||
|
CoSO4 x 7 H2O | 0.18 | g | ||
|
CaCl2 x 2 H2O | 0.10 | g | ||
|
ZnSO4 x 7 H2O | 0.18 | g | ||
|
CuSO4 x 5 H2O | 0.01 | g | ||
|
AlK(SO4)2 x 12 H2O | 0.02 | g | ||
|
H3BO3 | 0.01 | g | ||
|
Na2MoO4 x 2 H2O | 0.01 | g | ||
|
NiCl2 x 6 H2O | 0.03 | g | ||
|
Na2SeO3 x 5 H2O | 0.30 | mg | ||
|
Na2WO4 x 2 H2O | 0.40 | mg | ||
|
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. |