# 1264: SPIROCHAETA SP. MEDIUM


## Main sol. 1264

30	g	NaCl	
0.3	g	NH4Cl	
0.3	g	CaCl2 x 2 H2O	
3	g	MgCl2 x 2 H2O	
5	g	Starch (soluble)	
1	g	Yeast extract	
5	g	Peptone (Difco)	
10	ml	Phosphate buffer (10%, pH 7.4)	
1	ml	**Trace elements (Pfennig)**	
0.5	ml	Resazurin	
1000	ml	Distilled water	

1. Boil medium and cool under N2 stream on ice. Check pH (7.4). Autoclave aliquots and add the following sterile solutions (per 10 ml): 

0.1	ml	Na2S (3%)	
50	µl	Vitamine B12 (1 mg/10 ml)	


## Trace elements (Pfennig)

500	mg	EDTA	
200	mg	FeSO4 x 7 H2O	
10	mg	ZnSO4 x 7 H2O	
30	mg	H3BO3	
20	mg	CoCl2 x 6 H2O	
2	mg	NiCl2 x 6 H2O	
3	mg	Na2MoO4 x 2 H2O	
3	mg	MnCl2 x 4 H2O	
1	mg	CuCl2 x 2 H2O	
1000	ml	Distilled water