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DSMZ

CCAP 94 . SES:PJ

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Equipment needed:
Final pH: n.d.
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ml
Compound Amount Unit
Mail sol. C92 500 ml
Medium C76 500 ml
1 1:1 mixture
2 See separate recipes. Autoclave separately. Mix aseptically.

Mail sol. C92 #

1000
Compound Amount Unit
K2HPO4
(1 g/l stock solution) 		20 ml
MgSO4 x 7 H2O
(1 g/l stock solution) 		20 ml
KNO3
(10 g/l stock solution) 		20 ml
SE2 (Soil Extract 2) 100 ml
Deionized water 1000 ml
1 Make up to 1 litre with deionized water and autoclave at 15 psi for 15 minutes.
2 At the CCAP, SE1 is used for marine algae, SE2 for freshwater and terrestrial protozoa.

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.

SE1 (Soil Extract 1) #

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.

Mail sol. C76 #

1000
Compound Amount Unit
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) 		1 ml
Deionized water 1000 ml
1 Make up to 1 litre with deionized water. Autoclave at 15 psi for 15 minutes.

Stock solution (CaCl2/KCl) #

100
Compound Amount Unit
CaCl2 x 2 H2O 0.43 g
KCl 0.16 g