1291 .
CHTHONOMONAS MEDIUM
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Equipment needed:
Autoclave
Filter
Hungate tubes
Final pH:
5.0 - 5.3
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Gellan Gum
(Phytagel) |
0.50 | g | ||
|
|
(NH4)2SO4 | 4.00 | g | ||
|
|
K2HPO4 | 0.45 | g | ||
|
|
CaCl2 x 2 H2O | 0.15 | ml | ||
|
|
Trace element solution SL-11 | 1.00 | ml | ||
|
|
MgCl2 x 6 H2O | 1.00 | g | ||
|
|
Yeast extract | 0.01 | g | ||
|
|
Seven vitamins solution | 1.00 | ml | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Add gellan gum and boil to dissolve. Then add the remaining ingredients (except magnesium chloride, yeast extract and vitamins), adjust the pH value to 5.0 to 5.3 and fill the medium under an air atmosphere into tubes of the Hungate type or similar vials up to 30% their volume. After autoclaving add magnesium chloride, yeast extract and vitamins from steril stock solutions. Vitamins should be sterilized by filtration. | |||||
| 2 Note: Incubation with shaking may improve cell yield. | |||||
Trace element solution SL-11 #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Na2-EDTA x 2 H2O | 5.2 | g | ||
|
|
FeCl2 x 4 H2O | 1.5 | g | ||
|
|
ZnCl2 | 70.0 | mg | ||
|
|
MnCl2 x 4 H2O | 100.0 | mg | ||
|
|
H3BO3 | 6.0 | mg | ||
|
|
CoCl2 x 6 H2O | 190.0 | mg | ||
|
|
CuCl2 x 2 H2O | 2.0 | mg | ||
|
|
NiCl2 x 6 H2O | 24.0 | mg | ||
|
|
Na2MoO4 x 2 H2O | 36.0 | mg | ||
|
|
Distilled water | 1000.0 | ml | ||
| 1 Dissolve EDTA in 800 ml distilled water, adjust pH to 7 using 2 N NaOH and add ferrous chloride. After ferrous chloride has dissolved add other compounds. Finally adjust pH to 6.0 and bring volume to 1000 ml. | |||||
Seven vitamins solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Vitamin B12 | 100 | mg |
|
|
p-Aminobenzoic acid | 80 | mg |
|
|
D-(+)-biotin | 20 | mg |
|
|
Nicotinic acid | 200 | mg |
|
|
Calcium pantothenate | 100 | mg |
|
|
Pyridoxine hydrochloride | 300 | mg |
|
|
Thiamine-HCl x 2 H2O | 200 | mg |
|
|
Distilled water | 1000 | ml |