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DSMZ

CCAP 19 . BW/MASM

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Equipment needed:
Final pH: 8.0
Select a strain (optional):

ml
Compound Amount Unit
Tris 1 g
NaCl 30 g
MgSO4 x 7 H2O
(24.4 g/100 ml stock solution) 		10 ml
KCl
(6 g/100 ml stock solution) 		10 ml
NaNO3
(10 g/100 ml stock solution) 		10 ml
CaCl2 x 2 H2O
(3 g/100 ml stock solution) 		10 ml
KH2PO4
(0.5 g/100 ml stock solution) 		10 ml
NH4Cl
(2.67 g/100 ml stock solution) 		1 ml
Trace Elements (PIV) 6 ml
1 Once elements are dissolved autoclave at 15 psi for 15 minutes.
Vitamin B1 solution 1 ml
Vitamin B12 solution 1 ml
SE2 (Soil Extract 2) 30 ml
2 Make up to 1 litre with distilled water and adjust to pH 8.0 with1M NaOH or 1M HCl. Autoclave at 15 psi for 15 minutes.
3 For brackish organisms, take 15 g of NaCl instead of 30 g (BW/MASM).

Trace Elements (PIV) #

1000
1 Ensure elements are added in the following sequence:
Compound Amount Unit
Na2-EDTA 0.750 g
FeCl3 x 6 H2O 0.097 g
MnCl2 x 4 H2O 0.041 g
ZnCl2 0.005 g
CoCl2 x 6 H2O 0.002 g
Na2 MoO4 x 2 H2O 0.004 g
2 Once elements are dissolved autoclave at 15 psi for 15 minutes.

Vitamin B1 solution #

100
Compound Amount Unit
Thiamine hydrochloride
(Vitamin B1) 		0.12 g
1 Filter sterile

Vitamin B12 solution #

100
Compound Amount Unit
Cyanocobalamine
(Vitamin B12) 		0.1 g
1 Take 1 ml of this solution and add 99 ml Deionised water. Filter sterile.

SE2 (Soil Extract 2) #

1000
Compound Amount Unit
Soil
(air-dried) 		trace
Distilled water trace
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 handpicked for smaller invertebrates and roots. It should be turned periodically and picked over again. When dry sieve through a finer mesh (2-4 mm) and store in an airtight container away from light and heat.
3 Soil is prepared as above. Air-dried soil and twice its volume of supernatant distilled water are autoclaved together at 15 psi for 2 hours and left to cool. The supernatant is then decanted and then distributed to containers in volumes suitable for making up batches of media. The aliquots and their containers are autoclaved for an appropriate length of time (e.g. 1 litre or less for 15 minutes) and are then kept in a cool place (e.g. a refrigerator) until required.