Team:Calgary/Notebook/Protocols/Process5

From 2011.igem.org


Preparation of f2 Media for Algae

This protocol is used to prepare f2 media for algal growth.

Stock solutions

  • NaNO3 (150.0 g/L)
  • Trace metals (mixed to 750 mL to dissolve, and then brought up to 1 L): CuSO4.5H2O (19.6 mg/L), ZnSO4.7H2O (44.0 mg/L), CoCl2.6H2O (22.0 mg/L), MnCl2.4H2O (360.0 mg/L), Na2MoO4.2H2O (12.6 mg/L)
  • Na2SiO3.5H2O (22.7 g/L)
  • Fe citrate (both constituents added to the same 1 L: Ferric citrate (9.0g/L), Citric acid (9.0g/L)
  • Vitamin primary stocks: of Biotin (10.0 mg/100 mL) and vitamin B12 (10.0 mg/100 mL).
  • Vitamin working solutions (made fresh every three months) containing: 1.0mL primary stock Biotin, 1.0 mL Vitamin B12, 20.0 mg Thiamine HCL
  • NaH2PO4.2H2O (11.3 g/L)

    • Procedure

    1. Add 0.5 mL of each of stock solutions 1-5 (listed below) to 1 L of seawater
    2. Split into flasks and autoclave at 121 °C (15 PSI) for 15 minutes
    3. Prepare phosphate solution (see below) and autoclave dilute phosphate stock at 121°C (15PSI, 15 mins). After cooling, dispense aseptically with sterilised automatic dispenser. This component is autoclaved separately in order to prevent precipitation


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