SMMS: Difference between revisions
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*81.9 mL PEG 6000 (BDH 6.1% w/v in distilled water) | *81.9 mL PEG 6000 (BDH 6.1% w/v in distilled water) | ||
*2.5 mL Mg<sub>2</sub>SO<sub>4</sub>.7H<sub>2</sub>O - 5mM final | *2.5 mL Mg<sub>2</sub>SO<sub>4</sub>.7H<sub>2</sub>O - 5mM final | ||
*10 mL TES | *10 mL TES buffer (0.25M, pH 7.2) - 25mM final | ||
*1 mL NaH<sub>2</sub>PO<sub>4</sub> + K<sub>2</sub>HPO<sub>4</sub> (50mM each) - 1mM final | *1 mL NaH<sub>2</sub>PO<sub>4</sub> + K<sub>2</sub>HPO<sub>4</sub> (50mM each) - 1mM final | ||
*2 mL glucose (50% w/v) | *2 mL glucose (50% w/v) |
Latest revision as of 10:24, 19 July 2022
SMMS (Supplemented liquid minimal media, solid)
SMM is a solid supplemented minimal medium suitable for growth of many Streptomyces sp.
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 buffer (0.25M, pH 7.2) - 25mM final
- 1 mL 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)
Trace element solution: 0.1 g/L of the following:
- ZnSO4.7H2O
- FeSO4.7H2O
- MnCl2.4H2O
- CaCl2.6H2O
- NaCl
Make fresh every 2-4 weeks and filter-sterilise. Store at 4º
Instructions
- With the exception of the trace element solution each solution must be made up and autoclaved separately
- The solutions should be mixed in the given order and then made up to 1L with sterile distilled water
- Addition of 10 g/L agar
References
Strauch E, Takano E, Baylts H, Bibb M. The stringent response in Streptomyces coelicolor A3(2). Molecular Microbiology. 1991;5(2):289-298.