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DSMZ

DSMZ 1610 . METHYLOCEANIBACTER MEDIUM (NaCl-dANMS)

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Equipment needed:
Final pH: 7.8
Select a strain (optional):

ml
Compound Amount Unit
Solution 1 (50x salt stock) 20.0 ml
Solution 2 (100 mM Fe EDTA) 0.4 ml
Solution 3 (100 mM KH2PO4) 10.0 ml
Solution 4 (10 mM CuSO4 x 5 H2O) 0.1 ml
Solution 5 (30% NaCl) 100.0 ml
Solution 6 (100 mM NH4Cl) 20.0 ml
Solution 7 (100 mM KNO3) 20.0 ml
Solution 8 (100 mM Hepes) 50.0 ml
Solution 9 (100µM lanthanides) 1.0 ml
Trace element solution 1.0 ml
Distilled water 1000.0 ml
Methanol
(filter sterilized) 		10.0 ml
1 Dilute 20 ml of solution 1 to 700ml with distilled water.
2 Add 100mL of solution 5, 50 mL of solution 8, 20mL of solution 6 and 7 and 10 mL of solution 3.
3 Add 1ml trace element solution.
4 Add 0.4 mL solution 2.
5 Add 0.1 mL solution 4.
6 Add 1.0 mL solution 9.
7 Disolve and bring to 1L.
8 Adjust pH to 7.8 with HCl or NaOH. Autoclave medium.
9 After autoclaving add 10mL per liter of filter sterilized methanol.

Trace element solution #

1000
Compound Amount Unit
EDTA 0.50 g
FeSO4 x 7 H2O 0.20 g
ZnSO4 0.01 g
MnCl2 3.00 mg
H3BO3 0.03 g
CoCl2 x 6 H2O 0.02 g
CuCl2 x 5 H2O 0.01 g
NiCl2 x 6 H2O 0.02 g
Na2MoO4 x 2 H2O 2.00 mg
Distilled water 1000.00 ml
1 May be stored at 4°C in the dark

Solution 1 (50x salt stock) #

1000
Compound Amount Unit
MgSO4 x 6 H2O 10.0 g
CaCl2 x 2 H2O 1.5 g
Distilled water 1000.0 ml
1 Dissolve the ingredients listed above (in that order) in about 700 ml of distilled water, and then make up to 1 litre

Solution 2 (100 mM Fe EDTA) #

1000
Compound Amount Unit
Fe(III)-EDTA 36.05 g
Distilled water 1000.00 ml

Solution 3 (100 mM KH2PO4) #

1000
Compound Amount Unit
KH2PO4 13.6 g
Distilled water 1000.0 ml

Solution 4 (10 mM CuSO4 x 5 H2O) #

1000
Compound Amount Unit
CuSO4 x 5 H2O 2.5 g
Distilled water 1000.0 ml

Solution 5 (30% NaCl) #

1000
Compound Amount Unit
NaCl 30 g
Distilled water 1000 ml

Solution 6 (100 mM NH4Cl) #

1000
Compound Amount Unit
NH4Cl 5.3 g
Distilled water 1000.0 ml

Solution 7 (100 mM KNO3) #

1000
Compound Amount Unit
KNO3 10.1 g
Distilled water 1000.0 ml

Solution 8 (100 mM Hepes) #

1000
Compound Amount Unit
HEPES 23.8 g
Distilled water 1000.0 ml

Solution 9 (100µM lanthanides) #

1000
Compound Amount Unit
CeCl3 x 7 H2O 0.04 g
LaCl3 x 7 H2O 0.04 g
NdCl3 x 6 H2O 0.04 g
PrCl3 x H2O 0.02 g
Distilled water 1000.00 ml