# 889: ISS1 MEDIUM


## Main sol. 889

20	g	NaCl	
7	g	MgSO4 x 7 H2O	
5.5	g	MgCl2 x 6 H2O	
0.65	g	KCl	
0.1	g	NaBr	
0.3	g	NH4Cl	
0.2	g	K2HPO4	
0.5	g	CaCl2 x 2 H2O	
10	ml	**Modified Wolin's mineral solution II**	
0.2	g	Yeast extract	
5	g	Sulfur (powdered)	
1000	ml	Distilled water	

1. Adjust pH to 7.0 using H2SO4.
2. Prepare the medium under H2 gas and fill 20 ml medium into 100 ml serum bottles and seal with a rubber stopper. Steam/heat the medium in an almost boiling waterbath for 3 hours on each of 3 successive days to sterilize.
3. Add bicarbonate from a autoclaved and neutralized stock solution.

2	g/l	NaHCO3	

4. Check pH and adjust accordingly.
5. Give an atmosphere of H2/CO2/O2 (78/20/2) with an overpressure of two atmospheres.


## Modified Wolin's mineral solution II

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	
0.025	g	NiCl2 x 6 H2O	
0.3	mg	Na2SeO3 x 5 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.