# 640a: THERMOCLOSTRIDIUM CAENICOLA MEDIUM


## Main sol. 640a

0.9	g	NH4Cl	
0.9	g	NaCl	
0.4	g	MgCl2 x 6 H2O	
0.75	g	KH2PO4	
1.5	g	K2HPO4	
2	g	Trypticase peptone (BD BBL)	
1	g	Yeast extract (OXOID)	
1	ml	**Trace element solution SL-10**	
2.5	ml	FeCl3 x 6 H2O (0.1% w/v in 0.2 N HCl)	
0.5	ml	Sodium resazurin (0.1% w/v)	
2	g	NaHCO3	
1	g	Cellobiose	
0.5	g	L-Cysteine HCl x H2O	
1000	ml	Distilled water	

1. Dissolve ingredients except bicarbonate, cellobiose and cysteine. Sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic, then add bicarbonate and adjust pH to 6.8. Distribute medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add cellobiose and cysteine after autoclaving from anoxic stock solutions prepared under 100% N2 gas atmosphere. Cellobiose has to be sterilized by filtration. Adjust pH of the complete medium to 6.5, if necessary.


## Trace element solution SL-10

10	ml	HCl (25%)	
1.5	g	FeCl2 x 4 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 1000.00 ml.