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5x M9 Salts

  • Using premixed Difco 5x right now

M9 (for 1 L)

  1. Add 100 uL 1M CaCl2 to 750 mL autoclaved, warm water (see note)
  2. Add 2 mL 1M MgSo4
  3. Add 200 mL 5x M9 salts
  4. Add carbon source
  5. Add any other supplements
  6. Add autoclaved water to 1L
  • Make sure autoclaved water for diluting the M9, etc. is warm. M9 salts bottle suggests 40-50 deg C. I keep a bottle of autoclaved water for preparing this media in the 65 deg C incubator, so it's ready when I need it.

M9 Agar, 1L

  1. Add 200mL 5X M9 salts solution to ~800mL of distilled water
  2. Add 15g of agar
  3. Autoclave
  4. After autoclaving, swirl to mix evenly and "temper" at room temperature until you can place your hand on the flask for 2 second
  5. Add 2mL of 1M MgSO4, 0.1mL of 1M CaCl2 + carbon source of choice