29 .
E26 + Biotin:ANT
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Final pH:
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ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
E26 + biotin medium | 500 | ml | ||
|
|
Antia's medium (ANT) | 500 | ml | ||
| 1 E26 + biotin medium and Antia's medium are made up separately then mixed 1:1. | |||||
E26 + biotin medium #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Part 1 | 500 | ml | ||
|
|
Natural sea water
(filtered and autoclaved) |
500 | ml | ||
| 1 This medium is made up in 2 parts which are autoclaved separately and mixed aseptically, (1:1), when cool. This is to avoid precipitation. | |||||
| 2 Alternatively, use 17.5 g of "Ultramarine Synthetica" sea salts in 500 ml of distilled water. | |||||
Part 1 #
500
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
SE1 (Soil Extract 1) | 50.00 | ml | ||
|
|
KNO3 | 0.10 | g | ||
|
|
K2HPO4 | 0.01 | g | ||
|
|
MgSO4 x 7 H2O | 0.01 | g | ||
|
|
Cyanocobalamine | 0.10 | μg | ||
|
|
Thiamine HCl | 50.00 | μg | ||
|
|
Biotin | 0.10 | μg | ||
|
|
Distilled water | 500.00 | ml | ||
| 1 Distilled water to 500 ml. Autoclave | |||||
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 filtered through Whatman No 1 filter paper, 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. | |||||
Antia's medium (ANT) #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
KNO3 | 0.0500 | g | ||
|
|
NaH2PO4 x 2 H2O | 0.0078 | g | ||
|
|
Tris | 1.0000 | g | ||
|
|
Glycine | 0.3000 | g | ||
|
|
Trace metals stock solution (chelated) | 2.5000 | ml | ||
|
|
Thiamine HCl | 500.0000 | μg | ||
|
|
Cyanocobalamine | 2.0000 | μg | ||
|
|
Biotine | 1.0000 | μg | ||
|
|
Natural sea water
(filtered) |
800.0000 | ml | ||
| 1 Make up to 1 litre with distilled water and autoclave at 15 psi. Final pH should be 7.6 - 7.8. | |||||
Trace metals stock solution (chelated) #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Na2-EDTA x 2 H2O | 3.2400 | g | ||
|
|
FeCl3 x 6 H2O | 1.0800 | g | ||
|
|
MnSO4 x 4 H2O | 0.4500 | g | ||
|
|
ZnSO4 x 7 H2O | 0.2300 | g | ||
|
|
Na2MoO4 x 2 H2O | 0.0970 | g | ||
|
|
CuSO4 x 5 H2O | 0.0100 | g | ||
|
|
CoSO4 x 7 H2O | 0.0056 | g | ||
| 1 Make up to 1 litre with distilled water and adjust pH to 7.6 - 7.8 with dilute HCl or NaOH. Store frozen. | |||||