1002 .
YTPS MEDIUM
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Equipment needed:
Anaerobic workbench
Autoclave
Final pH:
7.3
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Yeast extract
(BD-Difco) |
5.00 | g | ||
|
|
Tryptone
(BD-Difco) |
5.00 | g | ||
|
|
Sodium pyruvate | 3.00 | g | ||
|
|
NaCl | 19.00 | g | ||
|
|
MgCl2 x 6 H2O | 9.00 | g | ||
|
|
MgSO4 x 7 H2O | 0.15 | g | ||
|
|
CaCl2 x 2 H2O | 0.30 | g | ||
|
|
KCl | 0.50 | g | ||
|
|
KH2PO4 | 0.42 | g | ||
|
|
(NH4)2SO4 | 0.10 | g | ||
|
|
NaBr | 0.05 | g | ||
|
|
SrCl2 x 6 H2O | 0.02 | g | ||
|
|
Na2SeO4 | 0.02 | g | ||
|
|
Wolfe's modified mineral elixir | 5.00 | ml | ||
|
|
Vitamin solution | 0.50 | ml | ||
|
|
PIPES | 6.05 | g | ||
|
|
L-Cysteine HCl x H2O | 0.50 | g | ||
|
|
Resazurin | 1.00 | mg | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Adjust pH to 7.3. | |||||
| 2 Mix ingredients, except L-cysteine x HCl x H2O, and bring to the boil while gassing with N2-H2 (95:5) gas mixture. When the color turns from blue to reddish pink, add L-cysteine x HCl x H2O during continuous gassing and boiling. Anaerobically dispense the medium in tubes under a N2-H2 (95:5) atmosphere. Stopper the tubes and autoclave. | |||||
Wolfe's modified mineral elixir #
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 | ||
|
|
Co(NO3)2 x 6 H2O | 0.10 | g | ||
|
|
CaCl2 | 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 | ||
|
|
Na2SeO3 | 1.00 | mg | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH. Then proceed to add minerals in the order listed. | |||||
Vitamin solution #
500
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Biotin | 20 | mg | ||
|
|
p-Aminobenzoic acid | 50 | mg | ||
|
|
Folic acid | 20 | mg | ||
|
|
Calcium pantothenate | 50 | mg | ||
|
|
Nicotinic acid | 50 | mg | ||
|
|
Vitamin B12 | 1 | mg | ||
|
|
Thiamine HCl | 5 | mg | ||
|
|
Pyridoxine hydrochloride | 100 | mg | ||
|
|
Thioctic acid | 50 | mg | ||
|
|
Riboflavin | 5 | mg | ||
|
|
Distilled water | 500 | ml | ||
| 1 Store at -20C or 4C in the absence of light. | |||||