Team:Calgary/Notebook/Protocols/Process5

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TITLE=Preparation of f2 Media for Algae|
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<p>
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This protocol is used to prepare f2 media for algal growth.</p>
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<h4> Stock solutions </h4>
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<li>NaNO<sub>3</sub> (150.0 g/L)</li>
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<li>Trace metals (mixed to 750 mL to dissolve, and then brought up to 1 L):  CuSO<sub>4</sub>.5H<sub>2</sub>O (19.6 mg/L), ZnSO<sub>4</sub>.7H<sub>2</sub>O (44.0 mg/L), CoCl<sub>2</sub>.6H<sub>2</sub>O (22.0 mg/L), MnCl<sub>2</sub>.4H<sub>2</sub>O (360.0 mg/L), Na<sub>2</sub>MoO<sub>4</sub>.2H<sub>2</sub>O (12.6 mg/L) </li>
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<li> Na<sub>2</sub>SiO<sub>3</sub>.5H<sub>2</sub>O (22.7 g/L) </li>
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<li>Fe citrate (both constituents added to the same 1 L: Ferric citrate (9.0g/L), Citric acid (9.0g/L)</li>
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<li>Vitamin primary stocks: of Biotin (10.0 mg/100 mL) and vitamin B12 (10.0 mg/100 mL).</li>
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<li>Vitamin working solutions (made fresh every three months) containing: 1.0mL primary stock Biotin, 1.0 mL Vitamin B12, 20.0 mg Thiamine HCL</li>
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<li> NaH<sub>2</sub>PO<sub>4</sub>.2H<sub>2</sub>O (11.3 g/L)</li>
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<h4> Procedure </h4>
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<li>Add 0.5 mL of each of stock solutions 1-5 (listed below) to 1 L of seawater</li>
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<li>Split into flasks and autoclave at 121 &deg;C (15 PSI) for 15 minutes </li>
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<li>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</li>
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Latest revision as of 04:05, 29 September 2011


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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