# 842: HALOANAEROBACTER SALINARIUS MEDIUM


## Main sol. 842

150	g	NaCl	
15	g	MgCl2 x 6 H2O	
3	g	KCl	
0.5	g	NH4Cl	
0.35	g	KH2PO4	
0.05	g	CaCl2 x 2 H2O	
0.1	g	Yeast extract	
0.5	g	Na2S x 9 H2O	
2	g	NaHCO3	
1.2	g	Glucose	
1	mg	Resazurin	
1	ml	**Trace element solution SL-12 B**	
1	ml	**Selenite-tungstate solution**	
1	ml	**Vitamin solution 7**	
997	ml	Distilled water	

1. Final pH 7.2 - 7.4
2. Prepare the medium without yeast extract, Na2S x 9 H2O, NaHCO3, glucose and vitamin solution (prepared as anaerobic, sterile stock solutions). Boil the medium and cool under anaerobic gas mixture of N2:CO2 (80:20). Dispense medium into Hungate tubes or serum bottles using the same gas mixture. Autoclave. Add the remaining components to the medium once it has cooled and check the final pH.


## Selenite-tungstate solution

6	mg	Na2SeO3 x 5 H2O	
8	mg	Na2WO4 x 2 H2O	
0.4	g	NaOH	
1000	ml	Distilled water	


## Vitamin solution 7

10	mg	Biotin	
50	mg	p-Aminobenzoic acid	
50	mg	Vitamin B12	
100	mg	Thiamine	
1000	ml	Distilled water	


## Trace element solution SL-12 B

3	g	Na2-EDTA	
1.1	g	FeSO4 x 7 H2O	
190	mg	CoCl2 x 6 H2O	
50	mg	MnCl2 x 2 H2O	
42	mg	ZnCl2	
24	mg	NiCl2 x 6 H2O	
18	mg	Na2MoO4 x 2 H2O	
300	mg	H3BO3	
2	mg	CuCl2 x 2 H2O	
1000	ml	Distilled water	

1. Adjust pH to 6.0.