# 1211: CALDISERICUM MEDIUM


## Main sol. 1211

0.05	g	K2HPO4	
0.09	g	KH2PO4	
0.25	g	NH4Cl	
0.25	g	MgSO4 x 7 H2O	
0.15	g	CaCl2 x 2 H2O	
5	g	Yeast extract	
1	ml	**Trace element solution**	
0.5	ml	Sodium resazurin (0.1% w/v)	
0.25	g	Na2CO3	
2.5	g	Na2S2O3 x 5 H2O	
2	ml	**Wolin's vitamin solution**	
0.25	g	L-Cysteine HCl x H2O	
0.25	g	Na2S x 9 H2O	
1000	ml	Distilled water	

1. Dissolve ingredients (except carbonate, thiosulfate, vitamins and reducing agents) and sparge medium with 80% N2 and 20% CO2 gas mixture 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 thiosulfate, vitamin solution, sulfide and cysteine 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. The pH of the complete medium should be at 6.5 - 6.8.


## Trace element solution

12.8	g	Nitrilotriacetic acid (NTA)	
1	g	FeCl2 x 4 H2O	
0.1	g	MnCl2 x 4 H2O	
0.03	g	CoCl2 x 6 H2O	
0.1	g	CaCl2 x 2 H2O	
0.1	g	ZnCl2	
0.02	g	CuCl2	
0.01	g	H3BO3	
0.03	g	Na2MoO4 x 2 H2O	
0.1	g	NiCl2 x 6 H2O	
1	g	NaCl	
0.03	g	Na2SeO3 x 5 H2O	
0.04	g	Na2WO4 x 2 H2O	
1000	ml	Distilled water	

1. First dissolve NTA in 200 ml of distilled water and adjust pH to 6.5 with KOH, then dissolve mineral salts. Finally adjust pH to 6.5 with KOH and make up to 1000.00 ml.


## 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