1525 .
KALLOTENUE MEDIUM
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Selected strain: Kallotenue papyrolyticum DSM 26889
This strain uses the standard medium without modifications.
Growth conditions: 2-3 days
55 °C
Equipment needed:
Autoclave
Filter
Final pH:
7.5
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
HEPES buffer
(SIGMA) |
3.57 | g | ||
|
|
NH4Cl | 27.00 | mg | ||
|
|
CaSO4 x 2 H2O | 60.00 | mg | ||
|
|
MgSO4 x 7 H2O | 0.10 | g | ||
|
|
NaCl | 8.00 | mg | ||
|
|
KNO3 | 0.10 | g | ||
|
|
NaNO3 | 0.69 | g | ||
|
|
Na2HPO4 | 0.11 | g | ||
|
|
Modified Wolin's mineral solution | 10.00 | ml | ||
|
|
D-Glucose | 2.00 | g | ||
|
|
Trypticase peptone
(BD BBL) |
1.00 | g | ||
|
|
Wolin's vitamin solution (10x) | 1.00 | ml | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Dissolve ingredients (except glucose, peptone and vitamins) and adjust pH to 8.1. Dispense medium under air atmosphere into culture vessels suitable for anaerobic cultivation (e.g., 10 ml medium per Balch-type tube), seal and autoclave. Add glucose, peptone and vitamins from sterile stock solutions sterilized by filtration. Adjust pH of complete medium to 7.5, if necessary. | |||||
Modified Wolin's mineral solution #
1000
| 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. | |||||
Wolin's vitamin solution (10x) #
1000
| 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 |