# 253: MEDIUM FOR ECTOTHIORHODOSPIRA


## Main sol. 253

0.5	g	KH2PO4	
0.05	g	CaCl2 x 2 H2O	
0.8	g	NH4Cl	
0.1	g	MgCl2 x 6 H2O	
180	g	NaCl	
20	g	Na2SO4	
6	g	Na2CO3	
1	ml	**Trace element solution (A)**	
0.5	g	Yeast extract	
1	g	Sodium succinate	
800	ml	Distilled water	

1. Adjust pH to 8.5.
2. Bubble with nitrogen gas and fill in tubes with a rubber septum under a stream of nitrogen gas. Autoclave at 121°C for 15 min. After sterilization add 1 ml/l filter-sterilized vitamin solution (B), 200 ml of filter-sterilized solution Bicarbonate solution (C) and 10 ml/l sterilised Na2S x 9H2O solution (D). pH 8.5

1	ml	**Vitamin solution (B)**	
200	ml	**Bicarbonate solution (C)**	
10	ml	**Na2S x 9H2O solution (D)**	


## Trace element solution (A)

1.8	g	FeCl2 x 4 H2O	
250	mg	CoCl2 x 6 H2O	
10	mg	NiCl2 x 6 H2O	
10	mg	CuCl2 x 2 H2O	
70	mg	MnCl2 x 4 H2O	
100	mg	ZnCl2	
500	mg	H3BO3	
30	mg	Na2MoO4 x 2 H2O	
10	mg	Na2SeO3 x 5 H2O	
1000	ml	Distilled water	

1. For dissolving adjust pH to about 3 with 1 N HCl.


## Vitamin solution (B)

10	mg	Biotin	
35	mg	Nicotinamide	
30	mg	Thiamine-HCl x 2 H2O	
20	mg	p-Aminobenzoic acid	
10	mg	Pyridoxal hydrochloride	
10	mg	Calcium pantothenate	
5	mg	Vitamin B12	
100	ml	Distilled water	


## Bicarbonate solution (C)

14	g	NaHCO3	
200	ml	Distilled water	


## Na2S x 9H2O solution (D)

10	g	Na2S x 9 H2O	
100	ml	Distilled water