687 .
MB MEDIUM WITH METHANOL
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Equipment needed:
Autoclave
Filter
Final pH:
n.d.
Select a strain (optional):
Solution A #
1001
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaCl | 10.0 | g | ||
|
|
Yeast extract | 2.0 | g | ||
|
|
Trypticase peptone | 2.0 | g | ||
|
|
NH4Cl | 1.0 | g | ||
|
|
MgCl2 x 6 H2O | 1.0 | g | ||
|
|
KCl | 0.5 | g | ||
|
|
CaCl2 x 2 H2O | 0.4 | g | ||
|
|
K2HPO4 | 0.4 | g | ||
|
|
Resazurin | 0.5 | mg | ||
|
|
Trace minerals | 10.0 | ml | ||
|
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Selenite-tungstate solution | 1.0 | ml | ||
|
|
NaHCO3 | 4.0 | g | ||
|
|
Distilled water | 920.0 | ml | ||
| 1 Mix components except NaHCO3, bring to a boil for several seconds and cool down under a stream of N2-CO2 (80:20). Add NaHCO3, mix thoroughly and distribute the medium into culture vessels under the same gas mixture. Seal with butyl rubber stoppers and autoclave. Finally, add per liter the following sterile solutions (autoclaved or *filter-sterilized): | |||||
|
|
Methanol solution (see below)
(*) |
50.0 | ml | ||
|
|
Trace vitamins (see Medium No. 197)
(*) |
10.0 | ml | ||
|
|
L-Cysteine HCl x H2O
(5%) |
5.0 | ml | ||
|
|
Na2S x 9 H2O
(5%) |
5.0 | ml | ||
Trace minerals #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Nitrilotriacetic acid | 1.50 | g | ||
|
|
MgSO4 x 7 H2O | 3.00 | g | ||
|
|
MnSO4 x n H2O | 0.50 | g | ||
|
|
NaCl | 1.00 | g | ||
|
|
FeSO4 x 7 H2O | 0.10 | g | ||
|
|
CoSO4 x 7 H2O | 0.10 | g | ||
|
|
CaCl2 x 2 H2O | 0.10 | g | ||
|
|
ZnSO4 x 7 H2O | 0.10 | g | ||
|
|
CuSO4 x 5 H2O | 0.01 | g | ||
|
|
AlK(SO4)2 | 0.01 | g | ||
|
|
H3BO3 | 0.01 | g | ||
|
|
Na2MoO4 x 2 H2O | 0.01 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH solution. Then proceed to add minerals. Adjust final pH to 7.0. | |||||
Selenite-tungstate solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
NaOH | 0.4 | g |
|
|
Na2SeO3 x 5 H2O | 6.0 | mg |
|
|
Na2WO4 x 2 H2O | 8.0 | mg |
|
|
Distilled water | 1000.0 | ml |