Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
KH2PO4 | 0.3 | g | ||
|
K2HPO4 | 0.3 | g | ||
|
NH4Cl | 0.5 | g | ||
|
KCl | 0.1 | g | ||
|
NaCl | 5.0 | g | ||
|
MgCl2 x 6 H2O | 0.5 | g | ||
|
CaCl2 x 2 H2O | 0.1 | g | ||
|
Yeast extract
(OXOID) |
5.0 | g | ||
|
Trypticase peptone
(BD BBL) |
5.0 | g | ||
|
Modified Wolin's mineral solution | 10.0 | ml | ||
|
Na2CO3 | 3.0 | g | ||
|
Na-crotonate solution (1 M) | 10.0 | ml | ||
|
Na2S x 9 H2O | 0.3 | g | ||
|
Distilled water | 1000.0 | ml | ||
1 Dissolve ingredients (except carbonate, crotonate and sulfide), adjust pH to 8.5 and sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic, then dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add crotonate and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile stock solution prepared under 80% N2 and 20% CO2 gas mixture. The crotonate solution should be sterilized by filtration. Adjust pH of the complete medium to 9.0, if necessary. |
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. |
Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
Crotonic acid
(ALDRICH 113018) |
86 | g | ||
|
NaOH
(10 N) |
100 | ml | ||
|
Distilled water | 900 | ml | ||
1 Dissolve crotonic acid in 800 ml distilled water, add around 100 ml of 10 N NaOH and adjust pH to around 7. Then add water to reach a volume of 1000 ml. Sterilize by filtration under 100% N2 gas atmosphere. |