Enrichment Solution:

NaNO3	4.7 g
Na2glycerophophate·5H2O	0.7 g
HEPES buffer	6.5 g
ES Fe Solution	325.0 ml
P-II Metal Solution	325.0 ml
Vitamin b12 Solution	3.0 ml
Biotin Vitamin Solution	3.0 ml
Thiamine Vitamin Solution	3.0 ml
Distilled water to	2.0 L

Final pH to 7.8; Add vitamins after autoclaving.

ES Fe Solution:

Fe(NH4)2(SO4)2·6H2O	1.4 g
Na2EDTA·2H2O	1.2 g
Distilled water to	2.0 L

P-II Metal Solution:

Na2EDTA·2H2O	0.1 g
H3BO3	0.114 g
FeCl3·6H2O	4.9 mg
MnSO4·H2O	16.4 mg
ZnSO4·7H2O	2.2 mg
CoCl2·6H2O	0.48 mg
Distilled water to	100 ml

Vitamin B₁₂ Solution:

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

Make HEPES solution at pH 7.8, then add Vitamin b₁₂ and filter-sterlize. Store dark in freezer.

Biotin Vitamin Solution:

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

Make HEPES solution at pH 7.8, then add biotin and filter-sterlize. Store dark in freezer.

Thiamine Vitamin Solution:

HEPES buffer	2.4 g
Thiamine	0.067 g
Distilled water to	200 ml

Make HEPES solution at pH 7.8, then add thiamine and filter-sterlize. Store dark in freezer.

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