990 .
MODIFIED MPOB MEDIUM
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Selected strain: Microbacter margulisiae DSM 27471
This strain uses the standard medium without modifications.
Equipment needed:
Anaerobic workbench
Filter
Final pH:
7.1
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Na2HPO4 | 0.53 | g | ||
|
|
KH2PO4 | 0.41 | g | ||
|
|
NH4Cl | 0.30 | g | ||
|
|
CaCl2 x 2 H2O | 0.11 | g | ||
|
|
MgCl2 x 6 H2O | 0.10 | g | ||
|
|
NaCl | 0.30 | g | ||
|
|
Trace element solution SL-10 | 1.00 | ml | ||
|
|
Yeast extract
(BD-Difco) |
0.40 | g | ||
|
|
Resazurin | 0.50 | mg | ||
|
|
Trace vitamins | 10.00 | ml | ||
|
|
Glucose | 1.80 | g | ||
|
|
L-Cysteine HCl x H2O | 0.50 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Prepare the medium anaerobically under a N2-CO2 (4:1, v/v) gas mixture. Mix ingredients except Trace vitamins. After autoclaving, anaerobically and aseptically add the filter-sterilized Trace vitamins to the medium. Check final pH to be 7.0 - 7.2. Pressurize the inoculated bottles to 70 kPa N2-CO2 (4:1, v/v). | |||||
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. | |||||
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 |