Modified COMBO Medium


NaNO3 solution (85.01 g/L)10 ml
CaCl2·2H2O solution (36.76 g/L)10 ml
MgSO4·7H2O solution (36.97 g/L)10 ml
NaHCO3 solution (12.6 g/L)10 ml
Na2SiO3·9 H2O solution (28.42 g/L)10 ml
K2HPO4 solution (8.71 g/L)1 ml
H3BO3 solution (24 g/L)10 ml
KCl solution (7.45 g/L)10 ml
Algal trace elements1 ml
Vitamin B12 solution1 ml
Biotin vitamin solution1 ml
Thiamine vitamin solution1 ml
CR1 soil1 cc
Deionized water to1 L

Do not autoclave. Filter sterilize. pH 7.8.

Algal trace element solution:
Na2EDTA·2H2O4.36 g
FeCl3·6H2O1 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 to1 L

Vitamin B12 solution:
HEPES buffer2.4 g
Vitamin B120.027 g
Deionized water to200 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 buffer2.4 g
Biotin0.005 g
Deionized water to200 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 buffer1.2 g
Thiamine0.11 g
Deionized water to100 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.


Modified from Guillard and Lorenzen (1972) WC medium for growth of cyanobacteria, cryptophytes, green algae, and diatoms at UTEX.



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