# 1049: SELENATE REDUCER MEDIUM


## Main sol. 1049

100	ml	**Solution A**	
1	ml	**Solution B**	
2.5	ml	**Solution C**	
1	ml	**Solution D**	
0.1	ml	**Solution E**	
0.5	ml	**Solution F**	
1	ml	**Solution G**	
1	ml	**Solution H**	

1. After cooling of medium A, add per 100 ml: 1.0 ml solution B, 2.5 ml solution C, 1.0 ml solution D, 0.1 ml solution E, 0.5 ml solution F, 1.0 ml solution G and 1.0 ml solution H.
2. Adjust pH to 7.0.
3. It is recommended to cultivate strain DSM 17993 on agar plates which may be prepared under aerobic conditions. Plates are then preincubated under anaerobic conditions prior to inoculation. They will stay pink coloured.
4. For liquid medium, prepare solution A, distribute to anaerobic cultivation vessels, heat to the boil and cool to room temperature while gassing with N2 : CO2 (80:20). Close vessels and autoclave. Prepare solutions B, C, D, E and H under anaerobic conditions and autoclave. Solutions F and H are filter sterilized and flushed with N2. Selenate (solution B) might be replaced by nitrate (solution: 1.00 g KNO3 in 10 ml).


## Solution A

1.12	g	DL-Na-lactate	
1.3	g	KCl	
0.2	g	KH2PO4	
23	g	NaCl	
0.5	g	NH4Cl	
0.1	g	CaCl2 x 2 H2O	
3	g	MgCl2 x 6 H2O	
0.5	mg	Resazurin	
15	g	Agar	
940	ml	Distilled water	


## Solution B

1.89	g	Na2SeO4	
10	ml	Distilled water	


## Solution C

1	g	NaHCO3	
10	ml	Distilled water	


## Solution D

0.1	g	Na2S x 9 H2O	
10	ml	Distilled water	


## Solution E

3	mg	Na2SeO3	
8	mg	Na2WO4	
100	ml	Distilled water	


## Solution F

1	ml	**Vitamin solution**	


## Solution G

1	ml	**Trace element solution SL-10**	
9	ml	Distilled water	


## Solution H

2	mg	1,4-Naphthoquinone	
0.5	mg	Hemin	
100	ml	Distilled water	


## 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.


## Vitamin solution

50	mg	Vitamin B12	
50	mg	Pantothenic acid	
50	mg	Riboflavin	
10	mg	Pyridoxamine hydrochloride	
20	mg	Biotin	
20	mg	Folic acid	
25	mg	Nicotinic acid	
25	mg	Nicotine amide	
50	mg	alpha-lipoic acid	
50	mg	p-Aminobenzoic acid	
50	mg	Thiamine-HCl x 2 H2O	
1000	ml	Distilled water	

1. Stir for some hours, filter sterilize the solution.