Lidstrom:MM1 recipe

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Trace Elements Recipe & Tips

• Total volume: 1L (adjust accordingly)
  • Solution #1 (500 mL ddH2O):
    • 4.4 g ZnSO4 (Heptahydrate)
    • 10 g EDTA*Na2 (Dihydrous)
  • Solution #2 (500 mL ddH2O):
    • 1.466 g CaCl2 (Dihydrate)
    • 1.012 g MnCl2 (Tetrahydrate)
    • 0.22 g (NH4)6Mo7O24 (Tetrahydrate)
    • 0.314 g CuSO4 (Pentahydrate)
    • 0.322 g CoCl2 (Hexahydrate)
    • 0.998 g FeSO4 (Heptahydrous)
• Procedure:

1. Prepare solution #1 mixture (in a 1L bottle) according to these criteria:
   1. Add EDTA*Na2 slowly, stirring with a stir bar while you do so.
   2. After EDTA*Na2 has been well dissolved (little to no cloudiness), add ZnSO4 scoop by scoop.
   3. For every 3 scoop, add 200 uL of 10 M KOH. This solution must not be incredibly acidic in order for each component to be soluble.
   4. You should end with a pH of 6. It would be optimal to make this solution with a pH probe attached.
2. Prepare solution #2 mixture (in a 1L bottle) and add a stir bar to assure that all components are dissolved well.
   1. Do not let this solution sit long before autoclaving. This solution should be light pink/orange by the time you combine it with solution #1.
3. Before autoclaving, pour equal amounts of solution #1 and #2 in smaller bottles (~100 mL total).
   1. At this point, if solution #2
4. Autoclave as soon as possible!

MM1 liquid recipe

• Total volume: 2L (adjust accordingly)
  • Solution #1 (1L ddH2O):
    • 5.06 g K2HPO4 (Anhydrous)
    • 5.18 g NaH2PO4 (Monohydrous)
  • Solution #2 (1L ddH2O):
    • 0.4 g MgSO4 (Heptahydrous)
    • 1 g (NH4)2SO4 (Anhydrous)
• Procedure:

1. Mix constituents of solution #1 and #2 separately in 2L bottles.
   1. Currently considering the effects of mixing #1 & #2 before autoclaving.
2. Organize tiny bottles to be filled with media post autoclave.
   1. Each bottle should contain ~150 mL of media.
3. Autoclave large bottles and tiny bottles.
4. Add 2 mL Visnaic trace elements into bottle #2.
5. Pour equal amounts of solution #1 and #2 into the smaller bottles.

MM1A recipe

• Total volume: 2L (adjust accordingly)
  • Solution #1 (1L ddH2O):
    • 5.06 g K2HPO4 (Anhydrous)
    • 5.18 g NaH2PO4 (Monohydrous)
  • Solution #2 (1L ddH2O):
    • 0.4 g MgSO4 (Heptahydrous)
    • 1 g (NH4)2SO4 (Anhydrous)
    • 30 g Bacto Agar
• Procedure:

1. Mix constituents of solution #1 and #2 separately in 2L bottles.
   1. Currently considering the effects of mixing #1 & #2 before autoclaving.
2. Organize tiny bottles to be filled with media post autoclave.
   1. Each bottle should contain ~150 mL of media.
3. Autoclave large bottles and tiny bottles.
4. Add 2 mL Visnaic trace elements into bottle #2.
5. Pour equal amounts of solution #1 and #2 into the smaller bottles.
6. Let cool until solidification for storage.
   1. If you wish to pour plates immediately after autoclaving, place bottles into 50C incubator before pouring into plates.