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DSMZ

CCAP 2 . AJS

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Equipment needed:
Final pH: 1.5
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ml
Compound Amount Unit
JM (Jaworski´s Medium) 970 ml
SE2 (Soil Extract 2) 30 ml
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) #

1000
Compound Amount Unit
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) 		1 ml
Deionized water 1000 ml
Agar , for solid medium
(Oxoid L11) 		15 g
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) #

660
Compound Amount Unit
Soil
(air­dried, sieved) 		105 g
Deionized water 660 ml
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 #

200
Compound Amount Unit
FeNa-EDTA 0.45 g
Na2-EDTA 0.45 g

Solution 2 #

200
Compound Amount Unit
H3BO3 0.496 g
MnCl2 x 4 H2O 0.278 g
(NH4)6Mo7O24 x 4 H2O 0.200 g

Solution 3 #

200
Compound Amount Unit
Cyanocobalamine 0.008 g
Thiamine HCl 0.008 g
Biotine 0.008 g