KH2PO4 1.0 g

KNO3 1.0 g

MgSO4 7 H2O 0.22 g

NaCl 30.0 g

Na2CO3 5.0 g

Trace element solution (see below) 1.0 ml

Vitamin B12 0.02 mg

Methanol 10.0 ml

Distilled water 1000.0 ml

Final pH 9.5

Trace elements solution:

Ferric citrate 30.0 mg

CaCl2 2 H2O 30.0 mg

MgCl2 4 H2O 5.0 mg

ZnSO4 7 H2O 5.0 mg

CuSO4 5 H2O 0.5 g

Distilled water 1000.0 ml

Prepare the medium without the Na2CO3, vitamin B12 and methanol. Basal medium, trace element and Na2CO3 autoclave separately at 121C 15 min. Add the vitamin B12 solution from a filter-sterilized stock solution and filter-sterilized methanol to the cooled medium. When preparing liquid media cool the mineral salts solution and Na2CO3 to room temperature before mixing. When preparing agar add 2.0 % agar to the mineral salt solution and autoclave. Cool the Na2CO3 stock solution and agar to 50-55C before mixing.


Updated 10.08.2021