# 1145: MS-MEDIUM (MODIFIED)


## Main sol. 1145

800	ml	Double distilled water (anaerobic)	
200	ml	**MS Buffer**	
5	g	Elemental sulphur	
2	g	NaS2O3 x 5 H2O	
7	g	MgSO4 x 7 H2O	
0.48	g	KCl	
0.4	g	CaCl2 x 2 H2O	
0.8	g	MgCl2 x 6 H2O	
20	ml	**Solution A**	
1.5	ml	**Solution B**	
10	ml	**Solution C - Trace mineral solution (Ferguson and Mah, 1983)**	

1. Each of the solutions (A, B, and C) is prepared with anaerobic double distilled water.
2. The medium is prepared anaerobically under nitrogen gas and dispensed into Hungate tubes or serum bottle sealed with rubber septa. The medium to gas phase ratio should be about 1:10 (v:v). 
Sterilise the medium by heating at 100° C for 3 hours on three consecutive days (e.g., in a water bath).
3. Add  4% O2 for microaerophilic conditions.


## MS Buffer

100	mM	NaOH	

1. 100 mM NaOH saturated with CO2


## Solution A

100	g	NH4Cl	
100	g	MgCl2 x 6 H2O	
40	g	CaCl2 x 2 H2O	
1000	ml	Distilled water	

1. Adjust to pH 4 with HCl


## Solution B

200	g	K2HPO4 x 3 H2O	
1000	ml	Distilled water	


## Solution C - Trace mineral solution (Ferguson and Mah, 1983)

500	mg	Na2-EDTA x 2 H2O	
30	mg	Na2WO4 x 2 H2O	
150	mg	CoCl2 x 6 H2O	
20	mg	CuCl2 x 2 H2O	
100	mg	MnCl2 x 4 H2O	
20	mg	NiSO4 x 6 H2O	
100	mg	FeSO4 x 7 H2O	
10	mg	H2SeO3	
100	mg	ZnCl2	
10	mg	H3BO3	
40	mg	AlCl3 x 6 H2O	
10	mg	Na2MoO4 x 2 H2O	
1000	ml	Distilled water	

1. Adjust pH to 3.0 with HCl