# 1243: PARADESULFITOBACTERIUM MEDIUM


## Main sol. 1243

13.7	g	Ferric citrate monohydrate	
1	g	NaCl	
0.4	g	MgCl2 x 6 H2O	
0.2	g	KH2PO4	
0.25	g	NH4Cl	
0.5	g	KCl	
0.15	g	CaCl2 x 2 H2O	
10	ml	**Modified Wolin's mineral solution**	
2.5	g	NaHCO3	
2.5	g	Na-pyruvate	
1	ml	**Wolin's vitamin solution (10x)**	
0.1	g	Na2S x 9 H2O	
1000	ml	Distilled water	

1. First dissolve ferric citrate by heating the water. After cooling to room temperature adjust the pH to 6.0 and dissolve the other ingredients (except bicarbonate, vitamins, pyruvate and sulfide). Sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Before inoculation add vitamins, pyruvate and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Stock solutions of pyruvate and vitamins should be sterilized by filtration. Adjust pH of complete medium to 6.8 - 7.2, if necessary.


## Modified Wolin's mineral solution

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	
0.03	g	NiCl2 x 6 H2O	
0.3	mg	Na2SeO3 x 5 H2O	
0.4	mg	Na2WO4 x 2 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.


## 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