# 1610: METHYLOCEANIBACTER MEDIUM (NaCl-dANMS)


## Main sol. 1610

20	ml	**Solution 1 (50x salt stock)**	
0.4	ml	**Solution 2 (100 mM Fe EDTA)**	
10	ml	**Solution 3 (100 mM KH2PO4)**	
0.1	ml	**Solution 4 (10 mM CuSO4 x 5 H2O)**	
100	ml	**Solution 5 (30% NaCl)**	
20	ml	**Solution 6 (100 mM NH4Cl)**	
20	ml	**Solution 7 (100 mM KNO3)**	
50	ml	**Solution 8 (100 mM Hepes)**	
1	ml	**Solution 9 (100µM lanthanides)**	
1	ml	**Trace element solution**	
1000	ml	Distilled water	
10	ml	Methanol (filter sterilized)	

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

0.5	g	EDTA	
0.2	g	FeSO4 x 7 H2O	
0.01	g	ZnSO4	
3	mg	MnCl2	
0.03	g	H3BO3	
0.02	g	CoCl2 x 6 H2O	
0.01	g	CuCl2 x 5 H2O	
0.02	g	NiCl2 x 6 H2O	
2	mg	Na2MoO4 x 2 H2O	
1000	ml	Distilled water	

1. May be stored at 4°C in the dark


## Solution 1 (50x salt stock)

10	g	MgSO4 x 6 H2O	
1.5	g	CaCl2 x 2 H2O	
1000	ml	Distilled water	

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)

36.05	g	Fe(III)-EDTA	
1000	ml	Distilled water	


## Solution 3 (100 mM KH2PO4)

13.6	g	KH2PO4	
1000	ml	Distilled water	


## Solution 4 (10 mM CuSO4 x 5 H2O)

2.5	g	CuSO4 x 5 H2O	
1000	ml	Distilled water	


## Solution 5 (30% NaCl)

30	g	NaCl	
1000	ml	Distilled water	


## Solution 6 (100 mM NH4Cl)

5.3	g	NH4Cl	
1000	ml	Distilled water	


## Solution 7 (100 mM KNO3)

10.1	g	KNO3	
1000	ml	Distilled water	


## Solution 8 (100 mM Hepes)

23.8	g	HEPES	
1000	ml	Distilled water	


## Solution 9 (100µM lanthanides)

0.04	g	CeCl3 x 7 H2O	
0.04	g	LaCl3 x 7 H2O	
0.04	g	NdCl3 x 6 H2O	
0.02	g	PrCl3 x H2O	
1000	ml	Distilled water