Bold 1NV Medium

NaNO₃ solution (25 g/L) 10 ml
P-IV metal solution 6 ml
CaCl₂ 2H₂O solution (2.5 g/L) 10 ml
MgSO₄ 7H₂O solution (7.5 g/L) 10 ml
K₂HPO₄ solution (7.5 g/L) 10 ml
KH₂PO₄ solution (17.5 g/L) 10 ml
NaCl solution (2.5 g/L) 10 ml
Vitamin B₁₂ solution 1 ml
Biotin vitamin solution 1 ml
Thiamine vitamin solution 1 ml
Deionized water to 1 L

Vitamins are added after autoclaving.

P-IV metal solution:
Na₂EDTA 2H₂O 0.75 g
FeCl₃ 6H₂O 0.097 g
MnCl₂ 4H₂O 0.041 g
ZnCl₂ 0.005 g
CoCl₂ 6H₂O 0.002 g
Na₂MoO₄ 2H₂O 0.004 g
Deionized water to 1 L

To approximately 950 ml of dH₂O, add ingredients in order listed stirring continuously. The EDTA should be fully dissoled before adding other components. Bring to 1 L with dH₂O and store at 4 C.

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.

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