Modified Bolds 3N Medium

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Modified Bolds 3N Medium Recipe

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

Directions

For 1 Liter Total (pH 6.2)

  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% agar medium add 15 g of agar into the flask; do not mix.
  3. Cover and autoclave medium.
  4. When cooled add vitamins.
    • For agar medium add vitamins, mix, and dispense before agar solidifies.
  5. Store at refrigerator temperature.

*Filter out soil particulates from Soilwater: GR+ before adding component to medium.


# 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    
8 Soilwater: GR+ Medium* 40 mL/L    
9 Vitamin B12 Solution 1 mL/L    
10 Biotin Vitamin Solution 1 mL/L    
11 Thiamine Vitamin Solution 1 mL/L