1723a .
Enriched Artificial Seawater Medium f/2 suppl. with Wheat Grains
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Equipment needed:
Final pH:
n.d.
Select a strain (optional):
Main sol. 1723a #
ml
| 1 Prepare 1l artificial sea water in milliQ water using: | |||||
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Artificial Sea Salt
, ~ 18 mS/cm = ~ 12 psu at 25°C
(hw Marinemix 21010, Wiegandt GmbH, Sterkenhofweg 13, 47807 Krefeld, Germany) |
12.4 | g/l | ||
| 2 Then add the following components: | |||||
|
|
NaNO3
(75 g/l stock solution) |
1.0 | ml/l | ||
|
|
NaH2PO4 x H2O
(5 g/l stock solution) |
1.0 | ml/l | ||
|
|
f/2 Trace Metal Mix | 1.0 | ml | ||
| 3 Autoclave for 20 min at 121°C. | |||||
| 4 After cooling, add the following filter sterilized (0.2 µm) components to complete the medium: | |||||
|
|
Vitamin 3 Mix | 1.0 | ml | ||
| 5 The strain is cultivated in 25 cm2 plastic tissue culture flasks with filter screw caps (T25, distributor: TPP, order number: 90026). The T25 flasks are routinely filled with 20 ml of culture. Each T25 culture is supplemented with a sterilized wheat grain as carbon source for ambient bacteria from the sampling site. | |||||
|
|
Wheat Grains
(sterilized by autoclaving at 121°C for 20 min) |
trace | |||
f/2 Trace Metal Mix #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Na2-EDTA x 2 H2O | 4.16 | g/l |
|
|
FeCl3 x 6 H2O | 3.15 | g/l |
|
|
MnCl2 x 4 H2O
(0.4 g/100 ml stock solution) |
45.00 | ml/l |
|
|
ZnSO4 x 7 H2O
(1.2 g/l stock solution) |
18.30 | ml/l |
|
|
CoCl2 x 6 H2O
(0.6 g/100 ml stock solution) |
1.70 | ml/l |
|
|
CuSO4 x 5 H2O
(1.25 g/100 ml stock solution) |
0.80 | ml/l |
|
|
Na2 MoO4 x 2 H2O
(0.27 g/100 ml stock solution) |
2.20 | ml/l |
Vitamin 3 Mix #
100
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Thiamine HCl
(1 g/100 ml stock solution) |
1.00 | ml |
|
|
D-Biotin
(0.05 g/100 ml stock solution) |
0.10 | ml |
|
|
Cyanocobalamin
(Vitamin B12; 0.2 g/l stock solution) |
0.25 | ml |