# 348: METHANOHALOPHILUS PORTUCALENSIS MEDIUM


## Main sol. 348

120	g	NaCl	
7	g	MgCl2 x 6 H2O	
6	g	MgSO4 x 7 H2O	
0.5	g	CaCl2 x 2 H2O	
0.4	g	K2HPO4 x 3 H2O	
1	g	NH4Cl	
3.8	g	KCl	
1	ml	**Trace element solution SL-10**	
1	ml	**Selenite-tungstate solution**	
2	g	Yeast extract (OXOID)	
2	g	Trypticase peptone (BD BBL)	
0.5	ml	Sodium resazurin (0.1% w/v)	
0.5	g	L-Cysteine HCl x H2O	
1.5	g	Na2CO3	
1.9	g	Trimethylamine-HCl	
0.25	g	Na2S x 9 H2O	
1000	ml	Distilled water	

1. Dissolve ingredients except cysteine, carbonate, trimethylamine and sulfide, bring medium to the boil, then cool to room temperature while sparging with 80% N2 and 20% CO2 gas mixture. Add cysteine and adjust pH to 6.5, then dispense under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Thereafter, add trimethylamine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Adjust pH of complete medium to 7.3 - 7.4, if necessary.


## Trace element solution SL-10

10	ml	HCl (25%)	
1.5	g	FeCl2 x 4 H2O	
70	mg	ZnCl2	
100	mg	MnCl2 x 4 H2O	
6	mg	H3BO3	
190	mg	CoCl2 x 6 H2O	
2	mg	CuCl2 x 2 H2O	
24	mg	NiCl2 x 6 H2O	
36	mg	Na2MoO4 x 2 H2O	
990	ml	Distilled water	

1. First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml.


## Selenite-tungstate solution

0.5	g	NaOH	
3	mg	Na2SeO3 x 5 H2O	
4	mg	Na2WO4 x 2 H2O	
1000	ml	Distilled water