DSMZ 645 . MIDDLEBROOK MEDIUM

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Selected strain: Mycobacterium neumannii DSM 43532

This strain uses the standard medium without modifications.


Growth conditions: 28 °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