# 702: HALOCELLA CELLULOLYTICA MEDIUM


## Main sol. 702

0.33	g	NH4Cl	
0.33	g	CaCl2	
0.33	g	MgCl2	
0.33	g	KCl	
0.33	g	KH2PO4	
150	g	NaCl	
2.5	g	NaHCO3	
0.5	g	Na2S x 9 H2O	
5	g	Glucose	
1	ml	**Trace element solution SL-10** (SL 10)	
10	ml	**Wolin's vitamin solution**	
2	mg	Resazurin	
2	g	Yeast extract	
3.3	g	MgCl2 x 6 H2O	
5	g	Cellulose (microcrystalline), alternative	
5	g	Cellobiose, alternative	
990	ml	Distilled water	

1. Prepare the medium without NaHCO3, Na2S, cellobiose or vitamins, boil and cool under a stream of nitrogen. Add the NaHCO3 and autoclave the medium. Na2S, cellobiose and vitamins are added from sterile, anaerobic stock solutions just before inoculating. When medium is prepared using microcrystalline cellulose instead of cellobiose as a substrate the microcrystalline cellulose must be added to the medium before autoclaving at the appropiate concentration. The final pH is 7.0.


## 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.


## 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