# 534: MODIFIED SME MEDIUM


## Main sol. 534

30	g	NaCl	
7	g	MgSO4 x 7 H2O	
5.5	g	MgCl2 x 6 H2O	
0.65	g	KCl	
0.1	g	NaBr	
0.15	g	NH4Cl	
0.15	g	K2HPO4	
0.5	g	CaCl2 x 2 H2O	
10	ml	**Trace element solution**	
1000	ml	Distilled water	

1. Sparge medium with 100% N2 gas mixture for 20 min to make it anoxic.
2. Adjust pH to 6.5-6.8 with 1N H2SO4.
3. Fill 10 ml of medium in serum bottles (100 ml) under a protective H2/CO2 atmosphere and tightly stopper.
4. Autoclave at 121 °C for 15 min.
5. after autoclaving add the following filter-sterilized solutions.

30	ml/l	**Solution A** (gassed with CO2), 0.3 ml/10ml	
30	ml/l	**Solution B** (gassed with N2), 0.3 ml/10ml	

6. apply xx bar H2/CO2 (80%/20%) + yy % O2 
(Note: strain specific conditions need to be applied)
7. For more details on the medium see the original description (Huber et al. System. Appl. Microbiol. 15:340-351 (1992)).


## Trace element solution

1000	ml	**Modified Wolin's mineral solution II**	

1. To 1000.0 ml trace element solution in medium 141 add:  

2	g	(NH4)2Ni(SO4)2	
10	mg	Na2WO4	
10	mg	Na2SeO4	


## Solution A

2	g	NaHCO3	
30	ml	Distilled water	

1. prepared under CO2 in an ice-bath, filter-sterilise


## Solution B

2	g	Na2S2O3 x 5 H2O	
30	ml	Distilled water	

1. Prepared under N2,autoclave 121°C, 15 min


## Modified Wolin's mineral solution II

1.5	g	Nitrilotriacetic acid	
3	g	MgSO4 x 7 H2O	
0.5	g	MnSO4 x H2O	
1	g	NaCl	
0.1	g	FeSO4 x 7 H2O	
0.18	g	CoSO4 x 7 H2O	
0.1	g	CaCl2 x 2 H2O	
0.18	g	ZnSO4 x 7 H2O	
0.01	g	CuSO4 x 5 H2O	
0.02	g	AlK(SO4)2 x 12 H2O	
0.01	g	H3BO3	
0.01	g	Na2MoO4 x 2 H2O	
0.025	g	NiCl2 x 6 H2O	
0.3	mg	Na2SeO3 x 5 H2O	
1000	ml	Distilled water	

1. First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Adjust final to pH 7.0 with KOH.