# C85: RPL/0.01%RPA


## Mail sol. C85

x	ml	**Mail sol. C82**	
x	ml	**0.01% RPA (Rye Grass­-Prescott Agar)**	

1. Biphasic (see separate recipes below and overleaf)


## Mail sol. C82

1	g	Rye grass (dried, milled)	
1000	ml	**Mail sol. C76**	

1. Bring PJ (Mail sol. C 76) to the boil then add rye grass.  Continue to boil for 5 minutes.  Allow to
cool then restore volume to 1 litre with deionized water.  Filter through GF/C paper 
and autoclave at 15 psi for 15 minutes.


## Mail sol. C76

1	ml	**Stock solution (CaCl2/KCl)**	
1	ml	K2HPO4 (0.51 g/100 ml stock solution)	
1	ml	MgSO4 x 7 H2O (0.28 g/100 ml stock solution)	
1000	ml	Deionized water	

1. Make up to 1 litre with deionized water. Autoclave at 15 psi for 15 minutes.


## Stock solution (CaCl2/KCl)

0.43	g	CaCl2 x 2 H2O	
0.16	g	KCl	


## 0.01% RPA (Rye Grass­-Prescott Agar)

0.1	g	Rye grass (dried, milled)	
1000	ml	**Mail sol. C76**	
15	g	Agar (Oxoid L11)	

1. Make up 1 litre of Rye Grass­Prescott Liquid (RPL -­  see separate recipe), but use 
only 0.1 g  of rye  grass.  Mix with 15 g of  Bacteriological Agar  (Oxoid L11).  Autoclave at 15 psi for 15 minutes.