# J1002: YTPS MEDIUM


## Main sol. J1002

5	g	Yeast extract (BD-Difco)	
5	g	Tryptone (BD-Difco)	
3	g	Sodium pyruvate	
19	g	NaCl	
9	g	MgCl2 x 6 H2O	
0.15	g	MgSO4 x 7 H2O	
0.3	g	CaCl2 x 2 H2O	
0.5	g	KCl	
0.42	g	KH2PO4	
0.1	g	(NH4)2SO4	
0.05	g	NaBr	
0.02	g	SrCl2 x 6 H2O	
0.02	g	Na2SeO4	
5	ml	**Wolfe's modified mineral elixir**	
0.5	ml	**Vitamin solution**	
6.05	g	PIPES	
0.5	g	L-Cysteine HCl x H2O	
1	mg	Resazurin	
1000	ml	Distilled water	

1. Adjust pH to 7.3.
2. Mix ingredients, except L-cysteine x HCl x H2O, and bring to the boil while gassing with N2-H2 (95:5) gas mixture. When the color turns from blue to reddish pink, add L-cysteine x HCl x H2O during continuous gassing and boiling. Anaerobically dispense the medium in tubes under a N2-H2 (95:5) atmosphere. Stopper the tubes and autoclave.


## Wolfe's modified mineral elixir

1.5	g	Nitrilotriacetic acid	
3	g	MgSO4 x 7 H2O	
0.5	g	MnSO4 x n H2O	
1	g	NaCl	
0.1	g	FeSO4 x 7 H2O	
0.1	g	Co(NO3)2 x 6 H2O	
0.1	g	CaCl2	
0.1	g	ZnSO4 x 7 H2O	
0.01	g	CuSO4 x 5 H2O	
0.01	g	AlK(SO4)2	
0.01	g	H3BO3	
0.01	g	Na2MoO4 x 2 H2O	
1	mg	Na2SeO3	
1000	ml	Distilled water	

1. Dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH. Then proceed to add minerals in the order listed.


## Vitamin solution

20	mg	Biotin	
50	mg	p-Aminobenzoic acid	
20	mg	Folic acid	
50	mg	Calcium pantothenate	
50	mg	Nicotinic acid	
1	mg	Vitamin B12	
5	mg	Thiamine HCl	
100	mg	Pyridoxine hydrochloride	
50	mg	Thioctic acid	
5	mg	Riboflavin	
500	ml	Distilled water	

1. Store at -20C or 4C in the absence of light.