104 .
PYG MEDIUM (modified)
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Selected strain: Isobaculum melis DSM 13760
This strain uses the standard medium without modifications.
Growth conditions: 37 °C
Equipment needed:
Autoclave
Filter
Final pH:
7.2
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Trypticase peptone | 5.0 | g | ||
|
|
Peptone | 5.0 | g | ||
|
|
Yeast extract | 10.0 | g | ||
|
|
Beef extract | 5.0 | g | ||
|
|
Glucose | 5.0 | g | ||
|
|
K2HPO4 | 2.0 | g | ||
|
|
Tween 80 | 1.0 | ml | ||
|
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Salt solution | 40.0 | ml | ||
|
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Resazurin | 1.0 | mg | ||
|
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Vitamin K1 solution | 0.2 | ml | ||
|
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Haemin solution | 10.0 | ml | ||
|
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L-Cysteine HCl x H2O | 0.5 | g | ||
|
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Distilled water | 950.0 | ml | ||
| 1 The vitamin K1, haemin solution and the cysteine are added after the medium has been boiled and cooled under CO2. Adjust pH to 7.2 using 8 N NaOH. Distribute under N2 and autoclave. | |||||
Salt solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
CaCl2 x 2 H2O | 0.25 | g |
|
|
MgSO4 x 7 H2O | 0.50 | g |
|
|
K2HPO4 | 1.00 | g |
|
|
KH2PO4 | 1.00 | g |
|
|
NaHCO3 | 10.00 | g |
|
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NaCl | 2.00 | g |
|
|
Distilled water | 1000.00 | ml |
Haemin solution #
100
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Haemin | 50 | mg | ||
|
|
NaOH
(1 N) |
1 | ml | ||
|
|
Distilled water | 100 | ml | ||
| 1 Dissolve 50 mg haemin in 1 ml 1 N NaOH; make up to 100 ml with distilled water and filter sterilize. Store refrigerated. | |||||
Vitamin K1 solution #
20
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Vitamin K1 | 0.1 | ml | ||
|
|
Ethanol
(95 %) |
20.0 | ml | ||
| 1 Dissolve 0.1 ml of vitamin K1 in 20 ml 95% ethanol and filter sterilize. Store refrigerated in a brown bottle. | |||||