Do not autoclave. Filter sterilize. pH 7.8.

Algal trace element solution:

Na₂EDTA 2H₂O 4.36 g
FeCl₃ 6H₂O 1 g
CuSO₄ 5 H₂O solution (0.01 g/ml) 1 ml
ZnSO₄ 7 H₂O solution (0.22 g/ml) 1 ml
CoCl₂ 6H₂O solution (0.12 g/ml) 1 ml
MnCl₂ 4H₂O solution (1.8 g/ml) 1 ml
Na₂MoO₄ 2H₂O solution (0.22 g/ml) 1 ml
H₂SeO₃ solution (0.016 g/ml) 1 ml
Na₃VO₄ solution (0.018 g/ml) 1 ml
Deionized water to 1 L

Vitamin B₁₂ solution:

HEPES buffer 2.4 g
Vitamin B₁₂ 0.027 g
Deionized water to 200 ml

Prepare 200 ml of HEPES buffer at pH 7.8 and fully dissolve vitamin. Filter-sterilize and store in dark at -20 C.

Biotin vitamin solution:

HEPES buffer 2.4 g
Biotin 0.005 g
Deionized water to 200 ml

Prepare 200 ml of HEPES buffer at pH 7.8 and fully dissolve vitamin. Filter-sterilize and store in dark at -20 C.

Thiamine vitamin solution:

HEPES buffer 1.2 g
Thiamine 0.11 g
Deionized water to 100 ml

Prepare 100 ml of HEPES buffer at pH 7.8 and fully dissolve vitamin. Filter-sterilize and store in dark at -20 C.

CR1 soil:
The CR1 soil is treated in batches by placing in a heat-resistant pan lined with aluminum foil. Fill the soil to a so depth of one-quarter inch and bake at 150 C for 2 h. Cool and cover the pan with aluminum foil and store in darkness at room temperature. Avoid excessive moisture during storage.

CR1 soil is not a well-defined medium, yet not all soil is suitable for culturing a broad range of algae. Soil used for CR1 originally came from the floodplain of the upper Rio de Flag, Flagstaff, Coconino Co., AZ. It is relatively low in organic content, pH circumneutral and is finely particulate in nature, consisting primarily of volcanic ash and weathered cinders and apparently contains a sufficient array of required trace elements and macronutrients.

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