Team:Valencia/BG-11Wl

From 2012.igem.org



BG-11 for Wildtype in medium liquid.


Reagents and Materials

  • Concentration BG-11 200X*:
    • 8 g/l de K2HPO4•3H2O.
    • 8 g/l de Na2CO3.
    • 15 g/l de MgSO4•7H2O.
    • 0,2 g/l de Na2EDTA.
    • 7,2 g/l de CaCl2•2H2O.
    • 1,2 g/l de ammonium Iron (III) citrate brown.
    • 1,2 g/l de citric acid anhydrous.

  • Concentration trace metal 1000X*:
    • 2,86 g/l de H3BO4.
    • 1,81 g/l de MnCl2•4H2O.
    • 222 mg/l de ZnSO4•7H2O.
    • 390 mg/l de Na2MoO4•2H2O.
    • 79 mg/l de CuSO4•5H20.
    • 49 mg/l de CoCl2•6H2O.


Protocol

  1. Add all the compounds for the BG-11 in an Erlenmeyer flask with 1 liter of distilled water and shake at room temperature until all salts are dissolved.
  2. Add all the compounds for the trace metal in an Erlenmeyer flask with 1 liter of distilled water and shake at room temperature until all salts are dissolved.
  3. In 1 liter of distilled water, put 1 Ml of trace metal and 5 ml of BG-11 and add 17’5 Mm of NaNO3.
  4. Autoclave at 121ºC during 20 minutes.
  5. Add 10 Mm of Hepes of NaOH to set the Ph=7’8 (1 M Hepes NaOH = 700 Ml distilled water, 283’3 g Hepes, ≈ 5’5 g NaOH).

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