# 331: ACETOBACTEROIDES GLYCINOPHILUS MEDIUM


## Main sol. 331

0.13	g	CaCl2 x 2 H2O	
1	g	NH4Cl	
0.2	g	MgCl2 x 6 H2O	
10	ml	**Trace element solution**	
5	ml	**Wolin's vitamin solution**	
1	mg	Resazurin	
0.5	g	Yeast extract	
1.5	g	Glycine	
3	g	KH2PO4	
5.8	g	Na2HPO4	
0.5	g	NaHCO3	
0.5	g	Na2S x 9 H2O	
1000	ml	Distilled water	

1. Adjust pH to 7.2 - 7.4 with NaOH.
2. Prepare the medium anaerobically under 100% nitrogen. Prepare concentrated solutions each of yeast extract, glycine, phosphates, NaHCO3 and Na2S separately and sterilize under nitrogen by autoclaving.


## Trace element solution

12.8	g	Nitrilotriacetic acid (NTA)	
0.2	g	FeCl2 x 4 H2O	
0.1	g	MnCl2 x 4 H2O	
0.17	g	CoCl2 x 6 H2O	
0.1	g	CaCl2 x 2 H2O	
0.1	g	ZnCl2	
0.02	g	CuCl2	
0.01	g	H3BO3	
0.01	g	Na2MoO4 x 2 H2O	
0.03	g	NiCl2 x 6 H2O	
1	g	NaCl	
0.03	g	Na2SeO3 x 5 H2O	
1000	ml	Distilled water	

1. First dissolve NTA in 200 ml of distilled water and adjust pH to 6.5 with KOH, then dissolve mineral salts. Finally adjust pH to 6.5 with KOH and make up to 1000.00 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