1668 .
MDM (modified dehority medium)
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Equipment needed:
Anaerobic workbench
Filter
Hungate tubes
Final pH:
6.8
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Yeast extract | 0.5 | g | ||
|
|
Tryptone | 0.5 | g | ||
|
|
Stock A: Salt solution 2x (50 ml stock per 100 ml medium) | 250.0 | ml | ||
|
|
Stock B: Salt solution 200x (0,5 ml stock per 100 ml medium) | 2.5 | ml | ||
|
|
Distilled water | 250.0 | ml | ||
|
|
L-Cysteine HCl | 1.0 | g/l | ||
| 1 To make medium anoxic boil it, cool under N2CO2 gas atmosphere, add 1 g/l L-cysteine hydrochloride and adjust pH to 6.8. Finally gas over with N2 . Dispense under N2CO2 into suitable vessels for anaerobic growth (serum bottles or Hungate tubes sealed with rubber stoppers) and sterilize by autoclaving. | |||||
Vitamin solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Biotin | 2.0 | mg | ||
|
|
Folic acid | 2.0 | mg | ||
|
|
Pyridoxine hydrochloride | 10.0 | mg | ||
|
|
Thiamine-HCl x 2 H2O | 5.0 | mg | ||
|
|
Riboflavin | 5.0 | mg | ||
|
|
Nicotinic acid | 5.0 | mg | ||
|
|
D-Calcium pantothenate | 5.0 | mg | ||
|
|
Vitamin B12 | 0.1 | mg | ||
|
|
p-Aminobenzoic acid | 5.0 | mg | ||
|
|
Lipoic acid | 5.0 | mg | ||
|
|
Distilled water | 1000.0 | ml | ||
| 1 Filter sterilize and make anoxic with N2. | |||||
Cellobiose (10%) #
100
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Distilled water | 100 | ml | ||
|
|
Cellobiose | 10 | g | ||
| 1 Mix 10 g cellobiose with 100 ml distilled water and filter sterilize, make anoxic with N2. | |||||
CM-Mix (only for DSM 104697) #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
D-Glucose | 4 | g | ||
|
|
Cellobiose | 1 | g | ||
|
|
Maltose | 1 | g | ||
|
|
Starch
(soluble) |
1 | g | ||
|
|
Distilled water | 1000 | ml | ||
| 1 Make anoxic with N2. | |||||
Vitamin K1 solution (only for DSM 104697) #
20
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Ethanol
(95 %) |
20.0 | ml | ||
|
|
Vitamin K1 | 0.1 | ml | ||
| 1 Dissolve 0.1 ml of vitamin K1 in 20 ml 95% ethanol and filter sterilize. Make anoxic with N2. Store refrigerated in a brown bottle. | |||||
Stock A: Salt solution 2x (50 ml stock per 100 ml medium) #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
K2HPO4 | 6804 | mg |
|
|
NaHCO3 | 6721 | mg |
|
|
NaCl | 4675 | mg |
|
|
NH4Cl | 1605 | mg |
|
|
CaCl2 | 89 | mg |
|
|
MgCl2 | 95 | mg |
|
|
FeSO4 x 7 H2O | 28 | mg |
|
|
Distilled water | 1000 | ml |
Stock B: Salt solution 200x (0,5 ml stock per 100 ml medium) #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
MgCl2 x 2 H2O | 971 | mg |
|
|
ZnCl2 | 273 | mg |
|
|
CoCl2 | 104 | mg |
|
|
Na2 MoO4 x 2 H2O | 48 | mg |
|
|
Na2SeO3 | 35 | mg |
|
|
NiCl2 x 6 H2O | 237 | mg |
|
|
Na2WO4 x 2 H2O | 66 | mg |
|
|
Distilled water | 1000 | ml |