Name: 3N-BBM+V (Bold-Basal Medium with 3-fold Nitrogen and Vitamins; modified)

Description: Freshwater algae

Stocks per litre
(1) NaNO3 - 25.0 g
(2) MgSO4.7H2O - 7.5 g
(3) NaCl - 0.5 g
(4) K2HPO4.3H2O - 7.5 g
(5) KH2PO4 - 17.5 g
(6) CaCl2.2H2O - 2.5 g

(7) Trace elements solution

      Add to 1 litre of distilled water 0.75 g Na2EDTA and the

      minerals below in exactly the following sequence:
      FeCl3.6H2O - 97.0 mg

      MnCl2.4H2O - 41.0 mg

      ZnCl2.6H2O - 5.0 mg

      CoCl2.6H2O - 2.0 mg

      Na2Mo4.2H2O - 4.0 mg

(8) Vitamin B1

      0.12 g Thiaminhydrochloride in 100 ml distilled water.  Filter sterile.

(9) Vitamin B12

      0.1 g Cyanocobalamin in 100 mg distilled water, take 1 ml of this solution

      and add 99 ml distilled water.  Filter sterile.

Medium

Stock solution 1 - 30.0 ml

Stock solutions 2 - 6, 10.0 ml each

Stock solution 7 - 6.0 ml
Stock solutions 8 - 9, 1.0 ml each

Method

Make up to 1 litre with deionized water.  Autoclave at 15 psi for 15 minutes.