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Modified Bold 3N Medium  

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

For 1 L 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.

  
#ComponentAmountStock Solution
Concentration
Final Concentration
1NaNO3 (Fisher BP360-500)30 mL/L10 g/400mL dH2O8.82 mM
2CaCl2·2H2O (Sigma C-3881)10 mL/L1 g/400mL dH2O0.17 mM
3MgSO4·7H2O (Sigma 230391)10 mL/L3 g/400mL dH2O0.3 mM
4K2HPO4 (Sigma P 3786)10 mL/L3 g/400mL dH2O0.43 mM
5KH2PO4 (Sigma P 0662)10 mL/L7 g/400mL dH2O1.29 mM
6NaCl (Fisher S271-500)10 mL/L1 g/400mL dH2O0.43 mM
7P-IV Metal Solution6 mL/L  
8Soilwater: GR+ Medium40 mL/L  
9Vitamin B121 mL/L  
10Biotin Vitamin Solution1 mL/L  
11Thiamine Vitamin Solution1 mL/L  

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