Media recipes
All solutions should be made up with distilled water. Media are usually prepared from stock solutions of macronutrients, trace metals, and vitamins which are added to a large proportion of the final volume of water in order to avoid precipitation. To prepare medium according to our recipes, first prepare the stock solutions in specified concentration (see column g / ...ml H2O). Then, add the specified volume of each stock solution (see column ml) into approximately 900 ml of distilled water. Finally, bring the total volume to 1000 ml with distilled water.
Media may be used as liquid or solidified by agar. In the list of strains, "ag." means that the strain is cultivated on the agarized medium, "liq." means it is cultivated on the liquid medium. To prepare agarized medium, add approximately 9-10 grams of agar to the 500 ml of liquid medium, and dissolve it in a steamer.
AFS - Acid Flagellates Solution
ASW - Artificial Sea Water
BBM - Bold´s Basal Medium with doubled nitrate
DAM - Dunaliella acidophila Medium
DY V - Modified DY IV Medium
SWM - Soil Water Medium
OGM - CAUP oligotrophic medium
Acid Flagellates Solution (AFS)
To approx. 900 ml of distilled water, add:
10 | NH4NO3 | |
10 | KH2PO4 | |
10 | MgSO4 . 7H2O | |
10 | CaCl2 | |
10 | Na-EDTA | |
10 | soil extract |
stock solutions B, H-Fe, H5, EDTA | |
vitamin B12 | 0.001 mg |
NaNO3 | 0.2 g |
Na2HPO4 | 0.03 g |
Seven Seas Utility Mix | 20 g |
soil extract | 50 ml |
Bold's Basal Medium (BBM) with doubled nitrate
To approx. 900 ml of distilled water, add:ml |
stock solution |
g / 400ml H2O |
20 | NaNO3 | 10 |
10 | CaCl2 . 2H2O | 1 |
10 | MgSO4 . 7H2O | 3 |
10 | K2HPO4 . 3H2O | 3 |
10 | KH2PO4 | 7 |
10 | NaCl | 1 |
1. Stock solution B | |
H3BO3 | 11.42 g |
dissolved in a 1000ml of distilled H2O |
2. Stock solution H-Fe | |
FeSO4 . 7H2O | 4.98 g |
conc. H2SO4 | 1 ml |
dissolved in a 1000ml of distilled H2O |
3. Stock solution H5 | |
ZnSO4 . 7H2O | 8.82 g |
MnCl2 . 4H2O | 1.44 g |
(NH4)6Mo7O24 . 4H2O | 0.88g |
CuSO4 . 5H2O | 1.57 g |
Co(NO3)2 . 6H2O | 0.49 g |
dissolved in a 1000ml of distilled H2O |
4. Stock solution EDTA | |
EDTA | 50 g |
KOH | 31 g |
dissolved in a 1000ml of distilled H2O |
Finally, bring the total volume to 1000 ml with distilled water.
Dunaliella acidophila Medium (DAM)
To approx. 950 ml of distilled water, add:MgSO4 . 7H2O | 25 g |
NH4NO3 | 0.25 g |
K2HPO4 | 0.075 g |
CaCl2 . 2H2O | 0.012 g |
peat extract | 5 ml |
stock solutions B, H-Fe, H5, EDTA | 1 ml each |
vitamin B12 | 0.0001 mg |
ml |
stock solution |
g / 100ml H2O |
1 | MgSO4 . 7H2O | |
1 | KCl | |
1 | NH4Cl | |
1 | NaNO3 | |
1 | β – glycerolphosphate | 0.216 |
1 | H3BO3 | |
1 | Na2EDTA | 0.7 |
1 | Na2SiO3 | |
1 | FeCl3 .6 H2O | 0.166 |
1 | CaCl2 | |
243 mg | HEPES |
Micronutrient solution |
|
MnCl2 . 4 H2O | 31.4 mg |
MnSO4 . H2O | 4.5 mg |
CoCl2 . 6 H2O | 1.46 mg |
Na2MoO4 . 2 H2O | 2.35 mg |
NH4VO3 | 0.13 mg |
Na2SeO3 | 0.27 mg |
dissolved in a 1000ml of distilled H2O |
Vitamin solution |
|
vitamin B12 | 0.005 g |
Thiamin HCl | 10 mg |
dissolved in a 1000ml of distilled H2O |
Soil Water Medium (SWM)
Place 1-2 cm of garden soil and cca 1 cm of sand in the bottom of a test tube (or Erlenmeyer flask). Fill up with distilled or deionized water. Autoclave for 30 min in two consecutive days.
CAUP oligotrophic Medium (OGM)
To approx. 900 ml of distilled water, add:ml |
stock solution |
g / 400ml H2O |
3.5 | NaNO3 | 10 |
2 | MgSO4 . 7H2O | 3 |
0.5 | KH2PO4 | 7 |
1 | CaSO4 | saturated solution |
0.2 | Stock solution B | |
5 | Stock solution H-Fe | |
0.2 | Stock solution H5 | |
1 | Stock solution EDTA | |
20 | soil extract | |
10 | Sphagnum extract |
Vitamin solution |
|
vitamin B12 | 0.005 g |
Thiamin HCl | 10 mg |
dissolved in a 1000ml of distilled H2O |
Note: This medium is actually not used in the collection.
Last modified on: | Copyright © Pavel Škaloud |