645 .
MIDDLEBROOK MEDIUM
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Selected strain: Mycobacterium longobardum DSM 45394
This strain uses the standard medium without modifications.
Growth conditions: 37 °C
Equipment needed:
Final pH:
n.d.
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Bacto Middlebrook 7H10 agar
(Difco 262710) |
19 | g | ||
|
|
Glycerol | 5 | ml | ||
|
|
Distilled water | 900 | ml | ||
| 1 Sterilize for 15 min by 121°C cool to 50 - 55°C. Add 100 ml Middlebrook OADC enrichment (Becton Dickinson 212240) under aseptic conditions and mix thoroughly. | |||||
| 2 If you can´t order the commercial media then prepare the media in this way: | |||||
Bacto Middlebrook 7H10 agar #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
(NH4)2SO4 | 0.500 | g | ||
|
|
KH2PO4 | 1.500 | g | ||
|
|
Na2HPO4 | 1.500 | g | ||
|
|
Sodium citrate | 0.400 | g | ||
|
|
MgSO4 | 0.025 | g | ||
|
|
CaCl2 | 0.500 | mg | ||
|
|
ZnSO4 | 1.000 | mg | ||
|
|
CuSO4 | 1.000 | mg | ||
|
|
L-Glutamic acid | 0.500 | g | ||
|
|
Ferric ammonium citrate | 0.040 | g | ||
|
|
Pyridoxine | 1.000 | mg | ||
|
|
Biotin | 0.500 | mg | ||
|
|
Malachite green | 0.250 | mg | ||
|
|
Agar
(Bacto ) |
15.000 | g | ||
|
|
Distilled water | 1000.000 | ml | ||
| 1 Final pH at 25°C 6.6 ± 0.2. | |||||
Bacto Middlebrook OADC enrichment #
100
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Oleic acid | 0.05 | g |
|
|
Bovine albumin
(albumin fraction V) |
5.00 | g |
|
|
Dextrose | 2.00 | g |
|
|
Catalase
(beef) |
4.00 | mg |
|
|
NaCl | 0.85 | g |
|
|
Distilled water | 100.00 | ml |