# 1663: BSM medium


## Main sol. 1663

3.62	g	NaCl	
0.26	g	K2HPO4	
0.08	g	(NH4)Cl	
0.85	g	NaNO3	
0.05	g	Na2SO4	
1	ml	**Trace element solution**	
10	ml	**Wolin's vitamin solution**	
1.12	g	Na-DL-lactate	
0.2	g	Yeast extract	
936.5	ml	Distilled water	
52.5	ml	NaHCO3 (8%)	

1. Dissolve ingredients (except vitamins and trace elements) in distilled water; boil for 1- 2 minutes and cool down to room temperature under 100% N2 gas, dispense in culture vessels and autoclave. After sterilization add NaHCO3, vitamins, and trace elements from sterile anoxic stock solutions prepared under 100% N2. Adjust pH to 8.2 and add sodium sulfide (Na2S) from a sterile anoxic solution to achieve a final concentration of 100 µM prior inoculation.


## Trace element solution

10	ml	HCl (25%; 7.7 M)	
1.5	g	FeCl2 x 4 H2O	
3	g	MgCl2 x 6 H2O	
0.1	g	CaCl2 x 2 H2O	
70	mg	ZnCl2	
100	mg	MnCl2 x 4 H2O	
6	mg	H3BO3	
190	mg	CoCl2 x 6 H2O	
2	mg	CuCl2 x 2 H2O	
24	mg	NiCl2 x 6 H2O	
36	mg	Na2MoO4 x 2 H2O	
990	ml	Distilled water	

1. First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.0 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