# 432: HALOBACTEROIDES ACETOETHYLICUS MEDIUM


## Main sol. 432

1000	ml	**Main sol. 144**	
100	g/l	NaCl	

1. Use medium no. 144 supplemented with 100 g/l of NaCl.
2. Sterilize glucose, yeast extract and sodium sulfide separately (autoclave under nitrogen atmosphere).


## Main sol. 144

0.9	g	NH4Cl	
0.9	g	NaCl	
0.4	g	MgCl2 x 6 H2O	
0.75	g	KH2PO4	
1.5	g	K2HPO4	
9	ml	**Trace element solution**	
3	ml	FeSO4 x 7 H2O (0.1% w/v in 0.1 N H2SO4)	
3	g	Yeast extract	
10	g	Trypticase peptone (BD BBL)	
0.5	ml	Sodium resazurin (0.1% w/v)	
5	ml	**Wolin's vitamin solution**	
5	g	D-Glucose	
1	g	Na2S x 9 H2O	
1000	ml	Distilled water	

1. Dissolve ingredients (except vitamins, glucose and sulfide), sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add glucose, vitamins (sterilized by filtration) and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Adjust pH of complete medium to 7.2 - 7.4.


## Wolin's vitamin solution

2	mg	Biotin	
2	mg	Folic acid	
10	mg	Pyridoxine hydrochloride	
5	mg	Thiamine HCl	
5	mg	Riboflavin	
5	mg	Nicotinic acid	
5	mg	Calcium D-(+)-pantothenate	
0.1	mg	Vitamin B12	
5	mg	p-Aminobenzoic acid	
5	mg	(DL)-alpha-Lipoic acid	
1000	ml	Distilled water