# 1076: SP4 MEDIUM


## Main sol. 1076

5	g	Tryptone	
2.65	g	Peptone	
1.75	g	PPLO broth	
375	ml	Distilled water	

1. Adjust pH to 7.4; autoclave at 121°C for 15 minutes.
2. Add aseptically: 

90	ml	Fetal bovine serum (heat-inactivated)	
25	ml	CMRL 1066 (10x concentrated; GIBCO)	
17.5	ml	Yeast extract (20%, autoclaved)	
5	ml	Yeast extract (20%, autoclaved)	
1	ml	Phenol red (1%, filter-sterilized)	
1.7	ml	L-Glutamine (200 mM; GIBCO)