M9 Minimal Medium



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tarix13.11.2017
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#10181

1M9 Minimal Medium
Per liter: Na2HPO4­·7H2O 12.8g

OR

Na2HPO4 (anhydrous) 6 g

KH2PO4 3 g

NaCl 0.5 g

NH4Cl 1 g
-add a carbon source (glucose, sodium gluconate, or glycerol) to 0.2% (v/v) final volume

-pH to 7.4 with NaOH

-Autoclave and then add sterile micronutrient components to a final concentration of:
Stock Concentration Micronutrient Final Concentration

1 M MgSO4 1 mM

1 M CaCl2 100 M

3 mM (NH4)6Mo7O24·4H2O 3x10-9 M

400 mM H3BO3 4x10-7 M

30 mM CoCl2 ·6H2O 3x10-8 M

10 mM CuSO4 ·5H2O 1x10-8 M

80 mM MnCl2 ·4H2O 8x10-8 M

10 mM ZnSO4 ·7H2O 1x10-8 M

5 mM FeSO4 ·7H2O§ 1x10-6 M




  • I recommend filter sterilization for all micronutrients

  • §FeSO4 must be made fresh and filter sterilized immediately before use. This solution is extremely prone to oxidation and precipitation, but may be frozen in aliquots.

Alternatively, make a 10X solution of the M9 salts, a 1000X mixed solution of the (NH4)6Mo7O24, H3BO3 , CoCl2 , CuSO4, MnCl2, and ZnSO4 and mix that with the appropriate FeSO4, MgSO4, CaCl2, and carbon source (with any antibiotics, arabinose, etc., etc.)


For 250mL of M9 solid medium (*autoclaved separately)
Component Volume Final concentration

1.5% (w/v) agar in MQ water* 200 mL 1% (w/v)

10X M9 salts* 25 mL 1X

20% (w/v) carbon source 2.5 mL 0.2% (w/v)

1000X micronutrients 0.25 mL 1X

1M MgSO4 0.25 mL 1 mM

1M CaCl2 0.025 mL 100 μM

5 mM FeSO 0.05 mL 1 μM



Milli-Q water* 21.925 mL

250mL
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