# C2: AJS


## Mail sol. C2

970	ml	**JM (Jaworski´s Medium)**	
30	ml	**SE2 (Soil Extract 2)**	

1. Acidified 97:3 mixture of JM and SE2
2. See separate recipes.  For approximately 1 litre of final medium, mix 970 ml JM
with 30 ml SE2.  Add 10 ml of concentrated H2SO4 to give a pH of approximately 
1.5.  Autoclave at 15 psi for 15 minutes.


## JM (Jaworski´s Medium)

1	ml	Ca(NO3)2 x 4 H2O (4.0 g/200 ml stock solution)	
1	ml	KH2PO4 (2.48 g/200 ml stock solution)	
1	ml	MgSO4 x 7 H2O (10.0 g/200 ml stock solution)	
1	ml	NaHCO3 (3.18 g/200 ml stock solution)	
1	ml	**Solution 1**	
1	ml	**Solution 2**	
1	ml	**Solution 3**	
1	ml	NaNO3 (16.0 g/200 ml stock solution)	
1	ml	Na2HPO4 x 12 H2O (7.2 g/200 ml stock solution)	
1000	ml	Deionized water	
15	g	Agar (Oxoid L11), for solid medium	

1. Make up to 1 litre with deionized water.  For agar, add 15.0 g per litre of Bacterial Agar (Oxoid L11).  Autoclave at 15 psi for 15 minutes.


## SE2 (Soil Extract 2)

105	g	Soil (air­dried, sieved)	
660	ml	Deionized water	

1. Site selection for a good soil is very important and for most purposes a soil from 
undisturbed deciduous woodland is best.  Sites to avoid are those showing obvious 
signs of man's activity and particular care should be taken to avoid areas where 
fertilizers, crop sprays or other toxic chemicals may have been used.
2. A rich loam with good crumb structure should be sought.  Stones, roots and larger 
invertebrates should be removed during an initial sieving through a 1 cm mesh.  The sieved soil should be spread to air dry and hand picked for smaller 
invertebrates and roots.  It should be turned periodically and picked over again.  When dry it may be sieved through a finer mesh (2­4 mm) or stored as it is prior to
use.
3. Soil is prepared as above.  105 g of air­dried sieved soil and 660 ml of deionized 
water are placed in a 1 litre bottle and autoclaved once at 15 psi for 15 minutes, 
then again after 24 hours.  The contents of the bottle are left to settle (usually for 
at least a week) and then the supernatant is decanted and filtered.  The final pH
should be 7.0 - 8.0.


## Solution 1

0.45	g	FeNa-EDTA	
0.45	g	Na2-EDTA	


## Solution 2

0.496	g	H3BO3	
0.278	g	MnCl2 x 4 H2O	
0.2	g	(NH4)6Mo7O24 x 4 H2O	


## Solution 3

0.008	g	Cyanocobalamine	
0.008	g	Thiamine HCl	
0.008	g	Biotine