Prov Medium

NaNO3 (75.0 g/L dH2O) 1.0 ml
NaH2PO4 H2O (5.0 g/L dH2O) 1.0 ml
Alkaline Soil Extract 15.0 ml
f/2 Trace Metal Solution 1.0 ml
f/2 Vitamin Solution 0.5 ml
Filtered seawater to 1.0 L

Autoclave after all additions.

Alkaline Soil Extract:
In a flask add two parts by volume distilled water (dH2O) to one part rich organic garden soil (containing no recent applications of chemical fertilizer or pesticides). Add 2-3 g NaOH/liter. Autoclave for 2 hours, and filter when cool. This concentrated extract is diluted 1:50 in dH2O for the final working stock. (Seawater may substituted for dH2O if the extract is to be used for marine species).

f/2 Trace Metal Solution:
FeCl3 6H2O 3.15 g
Na2EDTA 2H2O 4.36 g
CuSO4 5H2O (9.8 g/L dH2O) 1.0 ml
Na2MoO4 2H2O (6.3 g/L dH2O) 1.0 ml
ZnSO4 7H2O (22.0 g/L dH2O) 1.0 ml
CoCl2 6H2O (10.0 g/L dH2O) 1.0 ml
MnCl2 4H2O (180.0 g/L dH2O) 1.0 ml
Distilled water to 1.0 L


f/2 Vitamin Solution:
Vitamin B12 (1.0 g/L dH2O) 1.0 ml
Biotin (0.1 g/L dH2O) 10.0 ml
Thiamine HCl 200.0 mg
Distilled water to 1.0 L
Filter sterilize into plastic vials and store in refrigerator.

Note: Vitamin B12 and Biotin are obtained in a crystalline form. When preparing the Vitamin B12 Stock Solution allow for approximately 11% water of crystallization (For each 1.0 mg of Vitamin B12 add 0.89 ml dH2O). When preparing the Biotin Stock Solution allow for approximately 4% water of crystallization (For each 1.0 mg of Biotin add 9.6 ml dH2O).

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