# J438: RT2 MEDIUM


## Main sol. J438

1	g	NaCl	
0.1	g	MgCl2 x 6 H2O	
0.05	g	CaCl2 x 2 H2O	
0.25	g	NH4Cl	
0.1	g	KH2PO4	
0.5	g	KCl	
0.5	g	Yeast extract	
11.86	g	CAPSO	
4	g	Na2SO4	
1	ml	Trace element solution (see Medium No. 294)	
1	ml	**Selenite-tungstate solution**	
2.24	g	Sodium lactate	
1	mg	Resazurin	
0.1	g	Sodium dithionite	

1. Dissolve components except sodium lactate and sodium dithionite in 900 ml distilled water and adjust pH to 9.0 with NaOH. Bring to a boil, and cool down under a N2 gas stream. Distribute the medium into culture vessels (e.g., 9 ml of the medium in Hungate tubes) under N2, seal with butyl rubber stoppers and autoclave. Separately autoclave 0.2 M sodium lactate under a N2 atmosphere. After cooling, add 1/9 volume of the sodium lactate solution to the medium. Readjust pH to 9.0, if necessary. Prior to use, reduce the medium with a freshly prepared, filter-sterilize 1% sodium dithionite solution.


## Selenite-tungstate solution

0.4	g	NaOH	
6	mg	Na2SeO3 x 5 H2O	
8	mg	Na2WO4 x 2 H2O	
1000	ml	Distilled water