499 .
SULFATE-FREE SYNTROPHOMONUS MEDIUM
What is this page?
Equipment needed:
Autoclave
Filter
Final pH:
7.2
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Solution A | 916 | ml | ||
|
|
Solution B | 70 | ml | ||
|
|
Solution C | 10 | ml | ||
|
|
Solution D | 10 | ml | ||
| 1 Mix ingredients of Solution A, adjust pH to 7.2, bring to a boil for a few minutes and cool down to room temperature under a stream of N2-CO2 (4:1, v/v) gas mixture. Distribute the solution into culture vessels under the same gas mixture, seal with butyl rubber stoppers and autoclave. Filter-sterilize Solution B and equilibrate by gassing with the same gas mixture for at least 15 min. Autoclave Solutions C and D under a N2 atmosphere. Add appropriate amounts of Solutions B, C and D to Solution A to complete the medium. | |||||
Solution A #
916
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Mineral solution | 50.0 | ml |
|
|
Trace element solution SL-10 | 1.0 | ml |
|
|
Rumen fluid
, clarified
(see Medium No. 266) |
50.0 | ml |
|
|
Trypticase peptone
(BD-BBL) |
1.0 | g |
|
|
Vitamin solution | 5.0 | ml |
|
|
Crotonic acid | 1.7 | g |
|
|
Resazurin | 1.0 | mg |
|
|
Distilled water | 810.0 | ml |
Mineral solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
KH2PO4 | 10.0 | g |
|
|
MgCl2 x 6 H2O | 6.6 | g |
|
|
NaCl | 8.0 | g |
|
|
NH4Cl | 8.0 | g |
|
|
CaCl2 x 2 H2O | 1.0 | g |
|
|
Distilled water | 1000.0 | ml |
Vitamin solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 0.25 | mg |
|
|
Nicotinic acid | 2.50 | mg |
|
|
Thiamine HCl | 1.25 | mg |
|
|
p-Aminobenzoic acid | 1.25 | mg |
|
|
Pantothenic acid | 0.62 | mg |
|
|
Pyridoxine hydrochloride | 6.20 | mg |
|
|
Distilled water | 1000.00 | ml |
Trace element solution SL-10 #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
HCl
(25%, 7.7 M) |
10.0 | ml | ||
|
|
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 | 990.0 | ml | ||
| 1 Dissolve FeCl2 x 4H2O first in HCl. Then dilute in distilled water and add the remaining salts. | |||||