1051 .
ENDOMICROBIUM PROAVITUM RSA MEDIUM
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Equipment needed:
Anaerobic workbench
Autoclave
Filter
Final pH:
7.2
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaCl | 1.0 | g | ||
| 1 Adjust pH to 7.1 - 7.2. Distribute the medium into culture vessels under the same gas mixture and seal with butyl rubber stoppers. Finally, add per liter the following solution (filter-sterilized): | |||||
|
|
KCl | 0.5 | g | ||
|
|
MgCl2 x 6 H2O | 0.4 | g | ||
|
|
CaCl2 x 2 H2O | 0.1 | g | ||
|
|
NH4Cl | 0.3 | g | ||
|
|
KH2PO4 | 0.2 | g | ||
|
|
Resazurin | 0.4 | mg | ||
|
|
Distilled water | 945.0 | ml | ||
| 2 Mix components thoroughly and autoclave. Cool medium under a N2-CO2 (4:1, v/v) gas stream, and add the following solutions (autoclaved or *filter-sterilized) anaerobically and aseptically: | |||||
|
|
NaHCO3
(*, 8%) |
30.0 | ml | ||
|
|
Glucose
(*, 1 M) |
5.0 | ml | ||
|
|
L-Cysteine HCl x H2O
(0.1 M) |
10.0 | ml | ||
|
|
Yeast extract
(*, 10%) |
2.0 | ml | ||
|
|
Casamino acids
(*, 10%) |
2.0 | ml | ||
|
|
Trace element solution | 1.0 | ml | ||
|
|
FeCl2 solution | 1.0 | ml | ||
|
|
Selenite-tungstate solution | 1.0 | ml | ||
|
|
Trace vitamins solution (see Medium No. 284)
(*) |
1.0 | ml | ||
|
|
Fatty acid mixture | 1.0 | ml | ||
|
|
Aromatic fatty acids solution | 1.0 | ml | ||
|
|
Dithiothreitol
(250 mM) |
4.0 | ml | ||
Aromatic fatty acids solution #
100
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Phenyl acetic acid | 68 | mg | ||
|
|
Phenyl propionic acid | 75 | mg | ||
|
|
4-Hydroxyphenyl acetic acid | 76 | mg | ||
|
|
3-Indolyl acetic acid | 88 | mg | ||
|
|
NaOH
(20 mM) |
100 | ml | ||
| 1 Autoclave and store under a N2 atmosphere. | |||||
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 |
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 |
Selenite-tungstate solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
NaOH | 0.4 | g |
|
|
Na2SeO3 x 5 H2O | 6.0 | mg |
|
|
Na2WO4 x 2 H2O | 8.0 | mg |
|
|
Distilled water | 1000.0 | ml |
Fatty acid mixture #
20
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Valeric acid | 0.5 | g | ||
|
|
Isovaleric acid | 0.5 | g | ||
|
|
alpha-Methylbutyric acid | 0.5 | g | ||
|
|
Isobutyric acid | 0.5 | g | ||
|
|
Distilled water | 20.0 | ml | ||
| 1 Adjust pH to 7.5 with concentrated NaOH solution. | |||||