Bold 3N Medium



Bold 3N Medium Recipe

Modification of Bold's recipe. General purpose freshwater medium used for xenic cultures, especially blue-greens and reds.

Directions

For 1 L Total

  1. To approximately 850 mL of dH2O, add each of the components in the order specified (except vitamins) while stirring continuously.
  2. Bring the total volume to 1 L with dH2O.
    • *For 1.5% (w/v) agar medium, add 15 g of agar into the flaskl; do not mix.
  3. Cover and autoclave medium.
  4. When cooled add Vitamin B12.
    • *For agar medium add vitamin, mix, and dispense before agar solidifies.
  5. Store at refrigerator temperature.

# Component Amount Stock Solution Concentration Final Concentration
1 NaNO3 (Fisher BP360-500) 30 mL/L 10 g/400mL dH2O 8.82 mM
2 CaCl2·2H2O (Sigma C-3881) 10 mL/L 1 g/400mL dH2O 0.17 mM
3 MgSO4·7H2O (Sigma 230391) 10 mL/L 3 g/400mL dH2O 0.3 mM
4 K2HPO4 (Sigma P 3786) 10 mL/L 3 g/400mL dH2O 0.43 mM
5 KH2PO4 (Sigma P 0662) 10 mL/L 7 g/400mL dH2O 1.29 mM
6 NaCl (Fisher S271-500) 10 mL/L 1 g/400mL dH2O 0.43 mM
7 P-IV Metal Solution 6 mL/L    
9 Soilwater: GR+ Medium 40 mL/L    
10 Vitamin B12 Solution 1 mL/L