# 979: PETROBACTER SUCCINIMANDENS MEDIUM


## Main sol. 979

0.3	g	K2HPO4	
0.3	g	KH2PO4	
0.2	g	MgCl2 x 6 H2O	
0.1	g	CaCl2 x 2 H2O	
0.1	g	KCl	
1	g	NaCl	
1	g	NH4Cl	
2.36	g	Succinic acid	
1	g	Yeast extract	
5	ml	**Wolin's vitamin solution**	
10	ml	**Modified Wolin's mineral solution II**	
1000	ml	Distilled water	

1. Prepare the medium without the vitamins.
2. Adjust pH to 7.0 with 10M KOH.
3. Autoclave at 121 °C for 20 min.
4. After cooling add vitamins from a filter sterilised stock solution.


## 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	


## 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.