Transcription of M9 mineral medium - Helmholtz Zentrum München
1 M9 mineral mediumFor 1 liter m9 mineral medium add to 867 ml sterile water:100 mlM9 salt solution (10X) mM20 ml20% %1 ml1 M MgSO4 MgSO41 ml1 M mM1 mlbiotin (1 mg/ml)biotin1 g1 mlthiamin (1 mg/ml)thiamin1 g10 mltrace elements solution (100X)trace elements1 XStock solutionsM9 salt solution (10X) g/LKH2PO430 g/LNaCl5 g/LNH4Cl5 g/LDissolve the salts in 800 ml water and adjust the pH to with NaOH. Add water to afinal volume of 1 L and autoclave for 15 min at 121 Glucose20% (w/v) glucose200 g/LFor 500 ml stock solution add 100 g glucose to 440 ml water.
2 Autoclave for 15 min at121 M MgSO41 M g/100 mlFor 100 ml stock solution dissolve g MgSO4-7H2O in 87 ml water. Autoclave for 15min at 121 M CaCl21 M g/100 mlFor 100 ml stock solution dissolve 14,70 g CaCl2-2H2O in ml water. Autoclave for 15min at 121 (1 mg/ml)biotin (1mg/ml)50 mg/50 mlFor 50 ml stock solution dissolve 50 mg biotin in 45 ml water. Add small aliquots of 1 NNaOH until the biotin has dissolved. Add water to a final volume of 50 ml. Sterilize thesolution over a M filter. Prepare 1 ml aliquots and store at -20 (1 mg/ml)thiamin-HCl (1mg/ml)50 mg/50 mlFor 50 ml stock solution dissolve 50 mg thiamin-HCl in 45 ml water.
3 Add water to a finalvolume of 50 ml. Sterilize the solution over a m filter. Prepare 1 ml aliquots andstore at -20 trace elements solutionEDTA5 g mMZnCl284 mMCuCl2-2H2O13 mg/L76 MCoCl2-2H2O10 mg/L42 MH3BO310 mg/L162 MDissolve 5 g EDTA in 800 ml water and adjust the pH to with NaOH. Then add theother components in the quantities mentioned below and add water to a final volume of 1L. Sterilize the solution over a m mgFeCl3 (anhydrous)84 mgZnCl2765 M g/100 ml210 M g/100 M g/100 l1 M g/100 mlArie Geerlof - Helmholtz Center Munich15 May 2010
