Spizizen minimal medium

• 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!