Spizizen minimal medium

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  • Spizizen medium (SM) is a popular minimal medium for the cultivation of B. subtilis.

SM medium

  • 200 ml 5x SM base medium
  • 10 ml 100x Trace element solution
  • 10 ml 100x Fe-Citrate-Solution
  • 10 ml Glucose (50 %)
  • 10 ml Tryptophan (5 mg/ml) for B. subtilis 168
  • Add N-source solution to final concentration of 1 %
  • Ad 1,000 ml with deionized H2O

5x SM base medium

  • 175 g K2HPO4
  • 75 g KH2PO4
  • 12.5 g Na3-Citrate ∙ 2 H2O
  • 2.5 g MgSO4 ∙ 7 H2O
  • Ad 2,000 ml with deionized H2O

N-source solutions

  • Glutamine 4 % (do not autoclave)
  • Glutamate 40 %
  • (NH4)2 SO4, final concentration 0.2%

100x Trace element solution

  • 0.55 g CaCl2 / 0.735 g CaCl2 ∙ 2 H2O
  • 0.1 g MnCl2 ∙ 4 H2O
  • 0.17 g ZnCl2
  • 0.033 g CuCl2 ∙ 2 H2O / 0.043 g CuCl2 ∙ 6 H2O
  • 0.06 g CoCl2 ∙ 6 H2O
  • 0.06 g Na2MoO4 ∙ 2 H2O
  • Ad 1,000 ml with deionized H2O
  • Do not autoclave, use sterile filtration!

100x Fe-Citrate-Solution

  • 0.0135 g FeCl3 ∙ 6 H2O
  • 0.1 g Na3-Citrate ∙ 3 H2O
  • Ad 100 ml with deionized H2O
  • Do not autoclave, use sterile filtration!