731 .
SPOROMUSA MEDIUM
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Equipment needed:
Anaerobic workbench
Autoclave
Filter
Final pH:
7.0
Select a strain (optional):
Solution A #
863
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NH4Cl | 0.50 | g | ||
|
|
MgSO4 x 7 H2O | 0.50 | g | ||
|
|
CaCl2 x 2 H2O | 0.25 | g | ||
|
|
NaCl | 2.25 | g | ||
|
|
Trace vitamins | 10.00 | ml | ||
|
|
FeCl2 solution | 1.00 | ml | ||
|
|
Trace element solution | 1.00 | ml | ||
|
|
Se/W solution | 1.00 | ml | ||
|
|
Betaine | 6.70 | g | ||
|
|
Yeast extract | 2.00 | g | ||
|
|
Casitone
(BD-Difco) |
2.00 | g | ||
|
|
Resazurin | 1.00 | mg | ||
|
|
Distilled water | 850.00 | ml | ||
| 1 Autoclave under a N2 gas atmosphere. | |||||
Solution B #
100
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
K2HPO4 | 0.348 | g | ||
|
|
KH2PO4 | 0.227 | g | ||
|
|
Distilled water | 100.000 | ml | ||
| 1 Autoclave under a N2 gas atmosphere. | |||||
Solution C #
70
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaHCO3 | 4 | g | ||
|
|
Distilled water | 50 | ml | ||
| 1 To complete the medium, combine Solutions A, B and C, and replace the gas phase with a N2-CO2 (4:1, v/v) gas mixture. Finally, add the following solutions from anaerobic stocks: | |||||
|
|
L-Cysteine HCl x H2O
(3%) |
10 | ml | ||
|
|
Na2S x 9 H2O
(3%) |
10 | ml | ||
| 2 Filter-sterilize and store under a N2-CO2 (4:1, v/v) atmosphere. | |||||
| 3 Adjust pH of the medium to 7.0, if necessary. | |||||
Trace vitamins #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 2.0 | mg |
|
|
Folic acid | 2.0 | mg |
|
|
Pyridoxine hydrochloride | 10.0 | mg |
|
|
Thiamine HCl | 5.0 | mg |
|
|
Riboflavin | 5.0 | mg |
|
|
Nicotinic acid | 5.0 | mg |
|
|
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 |
FeCl2 solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
HCl
(7.7 M, 25%) |
10.0 | ml |
|
|
FeCl2 x 4 H2O | 1.5 | g |
|
|
Distilled water | 990.0 | ml |
Trace element solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
ZnCl2 | 70 | mg |
|
|
MnCl2 x 4 H2O | 100 | mg |
|
|
H3BO3 | 6 | mg |
|
|
CoCl2 x 6 H2O | 190 | mg |
|
|
CuCl2 x 2 H2O | 2 | mg |
|
|
NiCl2 x 6 H2O | 24 | mg |
|
|
Na2MoO4 x 2 H2O | 36 | mg |
|
|
Distilled water | 1000 | ml |