# J1238: TANNER MOD2 MEDIUM


## Main sol. J1238

2	g	NaCl	
0.25	g	NH4Cl	
0.25	g	KCl	
0.25	g	KH2PO4	
0.5	g	MgSO4 x 7 H2O	
0.1	g	CaCl2 x 2 H2O	
2.09	g	MOPS	
0.5	g	Yeast extract	
10	ml	**SL-9 mineral solution**	
0.5	g	L-Cysteine HCl x H2O	
0.5	mg	Resazurin	
990	ml	Distilled water	

1. Mix components thoroughly and adjust pH 7.0. Dispense the medium into culture vessels under a N2 gas atmosphere, seal with butyl rubber stoppers and autoclave. After cooling, add per liter the following solutions from sterile anaerobic stocks (filter-sterilized): 

30	ml	Fructose (1 M)	
10	ml	**Trace vitamins**	
1	ml	**Vitamin B12 solution 0.05 g/L**	
1	ml	Coenzyme M (0.01%)	


## SL-9 mineral solution

2	g	Nitrilotriacetic acid	
1	g	MnSO4 x n H2O	
0.8	g	Fe(NH4)2(SO4)2 x 6 H2O	
0.2	g	CoCl2 x 6 H2O	
1	mg	ZnSO4 x 7 H2O	
20	mg	CuCl2 x 2 H2O	
20	mg	NiCl2 x 6 H2O	
20	mg	Na2MoO4 x 2 H2O	
20	mg	Na2SeO4	
20	mg	Na2WO4 x 2 H2O	
1000	ml	Distilled water	

1. Dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH. Then proceed to add minerals. Adjust final pH to 7.0.


## Trace vitamins

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 pantothenate	
0.1	mg	Vitamin B12	
5	mg	p-Aminobenzoic acid	
5	mg	Lipoic acid	
1000	ml	Distilled water	


## Vitamin B12 solution 0.05 g/L

5	mg	Vitamin B12	
100	ml	Distilled water