SMM

SMM (Supplemented liquid minimal media)

Strauch et al., 1991¹

Preparation

Per litre:

• 81.9 mL PEG 6000 (BDH 6.1% w/v in distilled water)
• 2.5 mL Mg₂SO₄.7H₂O - 5mM final
• 10 mL TES bugger (0.25M, pH 7.2) - 25mM final
• 1mL NaH₂PO₄ + K₂HPO₄ (50mM each) - 1mM final
• 2 mL glucose (50% w/v)
• 1 mL Antifoam 289 (Sigma A 5551)(1% w/v)
• 0.1 mL Trace element solution
• 1 mL Difco Casaminoacids (20% w/v)
• 100 mL Total
• 25 ml/L Glycine (0.5% final)

Instructions

1. Dissolve all of the ingredients except the agar and the glucose in the distilled water
2. Adjust the pH to 7.0-7.2
3. Dispense 200ml into flasks each containing 2g agar
4. Autoclave
5. After autoclaving, add 4 ml sterile 50% glucose solution (autoclaved separately or filter-sterilized) to each flask
6. Add any other growth factors required for auxotrophic mutants