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



Acid Flagellates Solution (AFS)
To approx. 900 ml of distilled water, add:

ml
stock solution
g /100ml H2O
10   NH4NO3
0.25
10   KH2PO4
0.15
10   MgSO4 . 7H2O
0.25
10   CaCl2
0.05
10   Na-EDTA
0.025
10   soil extract  

and

stock solutions B, H-Fe, H5, EDTA
1ml  each
vitamin B12 0.001 mg

Finally, bring the total volume to 1000 ml with distilled water. Adjust to pH 3.5 with 1M HCl  


Artificial Sea Water (ASW)

To 950 ml of distilled water, add:

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

Then, add 1 ml of each of following 4 stock solutions to 1000 ml of medium:

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

Finally, bring the total volume to 1000 ml with distilled water. Adjust to pH 1.5-1.7 with 1M H2SO4  


Modified DY V Medium

To aprox. 950 ml of distilled water, add:

ml
stock solution
g / 100ml H2O
1    MgSO4 . 7H2O
5
1    KCl
0.3
1    NH4Cl
0.268
1    NaNO3
2
1    β – glycerolphosphate
0.216
1    H3BO3
0.08
1    Na2EDTA
0.7
1    Na2SiO3
0.6
1    FeCl3 .6 H2O
0.166
1    CaCl2
7.5
243 mg    HEPES
-

Then, add 1 ml of following micronutrient solution:

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

Then, add 1 ml of following vitamin solution:

Vitamin solution
vitamin B12 0.005 g
Thiamin HCl 10 mg
dissolved in a 1000ml of distilled H2O

Finally, bring the total volume to 1000 ml with distilled water..  


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.





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Copyright © Pavel Škaloud