# 968b: MLMS-1 MEDIUM (ARSENATE)


## Main sol. 968b

0.1	g	(NH4)2SO4	
25	mg	MgSO4	
60	g	NaCl	
0.15	g	K2HPO4	
0.08	g	KH2PO4	
1	ml	**Trace element solution SL-10**	
1	ml	**Selenite-tungstate solution**	
0.5	ml	Sodium resazurin (0.1% w/v)	
10.6	g	Na2CO3	
4.2	g	NaHCO3	
3.1	g	Na2HAsO4 x 7 H2O	
1	ml	**Wolin's vitamin solution (10x)**	
0.82	g	Na2S x 9 H2O	
1000	ml	Distilled water	

1. Dissolve ingredients (except carbonates, arsenate, vitamins and sulfide), sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Add solid carbonate and hydrogencarbonate, adjust pH to 9.8, dispense medium under 100% N2 gas atmosphere into anoxic Hungate-type tubes or seruim vials and autoclave. Prior to inoculation, add arsenate, vitamins (sterilized by filtration) and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Adjust pH of complete medium to 9.8, 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	


## Wolin's vitamin solution (10x)

20	mg	Biotin	
20	mg	Folic acid	
100	mg	Pyridoxine hydrochloride	
50	mg	Thiamine HCl	
50	mg	Riboflavin	
50	mg	Nicotinic acid	
50	mg	Calcium D-(+)-pantothenate	
1	mg	Vitamin B12	
50	mg	p-Aminobenzoic acid	
50	mg	(DL)-alpha-Lipoic acid	
1000	ml	Distilled water