# J1181: MODIFIED CELLULOLYTIC HALOARCHAEA MEDIUM


## Main sol. J1181

833	ml	**MDS salt water**	
1	ml	**Trace element solution**	
0.1	g	Yeast extract (BD-Difco)	

1. Add components and bring volume to 958 ml with distilled water. For preparation of solid medium, add 20 g/L agar (if necessary, use washed agar: see Medium No. 574). Autoclave. After cooling, add the following solutions (autoclaved or *filter-sterilized) to the medium. 

5	ml	NH4Cl (1 M)	
2	ml	**Potassium phosphate buffer**	
5	ml	**Trace vitamins** (*)	
30	ml	Cellobiose (0.2 M)	

2. Readjust pH to 7.0 with a sterilized 10% Na2CO3 solution, if necessary.


## MDS salt water

240	g	NaCl	
30	g	MgCl2 x 6 H2O	
35	g	MgSO4 x 7 H2O	
7	g	KCl	
5	ml	CaCl2 (1 M)	

1. Add components to distilled water and bring volume to 1.0 L. Adjust pH to 7.5 with 1 M Tris base.


## Trace element solution

5	g	EDTA	
2	g	FeSO4 x 7 H2O	
0.1	g	ZnSO4 x 7 H2O	
0.03	g	MnCl2 x 4 H2O	
0.3	g	H3BO3	
0.2	g	CoCl2 x 6 H2O	
0.01	g	CuCl2 x 2 H2O	
0.02	g	NiCl2 x 6 H2O	
0.03	g	Na2MoO4 x 2 H2O	
1000	ml	Distilled water	

1. Adjust pH to 3.6.


## Potassium phosphate buffer

83.4	ml	K2HPO4 (1 M)	
16.6	ml	KH2PO4 (1 M)	

1. Combine the two solutions and check that pH is close to 7.5. Add an equal volume of distilled water.


## Trace vitamins

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 pantothenate	
0.1	mg	Vitamin B12	
5	mg	p-Aminobenzoic acid	
5	mg	Lipoic acid	
1000	ml	Distilled water