# 1533: MARINIFILUM MEDIUM (IRD MEDIUM)


## Main sol. 1533

0.3	g	KH2PO4	
0.3	g	K2HPO4	
1	g	NH4Cl	
25	g	NaCl	
0.1	g	KCl	
0.1	g	CaCl2 x 2 H2O	
0.5	g	Cysteine HCl	
1	g	Yeast extract (Oxoid)	
1	mL	**Trace element solution SL-7**	
1	mL	Resazurin (0.1%)	
1000	ml	Distilled water	

1. Add after autoclave 

20	ml/l	MgCl2 x 6 H2O (150.0 g/l)	
20	ml/l	NaHCO3 (10%)	
20	ml/l	Na2S x 9 H2O (2%)	
20	ml/l	Glucose (1M)	

2. pH 7.0
3. Prepare the medium without NaHCO3, Na2S x 9 H2O, MgCl2 x 6 H2O and glucose. Boil the medium in a water bath and cool under a stream of N2/CO2 (80/20). Add the NaHCO3 to the cooled medium (the pH should be 7.0) and dispense the medium into Hungate tubes or serum bottles under a stream of N2/CO2 (80/20). Seal the tubes under N2/CO2 (80/20) and autoclave. To the cooled medium at the appropriate amounts of Na2S x 9 H2O, MgCl2 x 6 H2O and glucose from sterile, anaerobic stock solutions.


## Trace element solution SL-7

1000	ml	Distilled water	
6.5	ml	HCl (25%)	
1.5	g	FeCl2 x 7 H2O	
70	mg	ZnCl2	
100	mg	MnCl2 x 4 H2O	
62	mg	H3BO3	
190	mg	CoCl2 x 6 H2O	
17	mg	CuCl2 x 2 H2O	
24	mg	NiCl2 x 6 H2O	
36	mg	Na2MoO4 x 2 H2O