SMM: Difference between revisions

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=SMM (Supplemented liquid minimal media)=
=SMM (Supplemented liquid minimal media)=
Strauch et al., 1991<sup>1</sup>
SMM is a supplemented minimal medium suitable for growth of many ''Streptomyces'' sp.


==Preparation==
==Preparation==
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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 bugger (0.25M, pH 7.2) - 25mM final
*10 mL TES buffer (0.25M, pH 7.2) - 25mM final
*1mL 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)
*1 mL Antifoam 289 (Sigma A 5551)(1% w/v)
*1 mL Antifoam 289 (Sigma A 5551)(1% w/v)
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*100 mL Total
*100 mL Total
*25 ml/L Glycine (0.5% final)
*25 ml/L Glycine (0.5% final)
Trace element solution:
0.1 g/L of the following:
*ZnSO<sub>4</sub>.7H<sub>2</sub>O
*FeSO<sub>4</sub>.7H<sub>2</sub>O
*MnCl<sub>2</sub>.4H<sub>2</sub>O
*CaCl<sub>2</sub>.6H<sub>2</sub>O
*NaCl
Make fresh every 2-4 weeks and filter-sterilise. Store at 4&ordm;




'''Instructions'''
'''Instructions'''
#Dissolve all of the ingredients except the agar and the glucose in the distilled water
#With the exception of the trace element solution each solution must be made up and autoclaved separately
#Adjust the pH to 7.0-7.2
#The solutions should be mixed in the given order and then made up to 1L with sterile distilled water
#Dispense 200ml into flasks each containing 2g agar
 
#Autoclave
==References==
#After autoclaving, add 4 ml sterile 50% glucose solution (autoclaved separately or filter-sterilized) to each flask
Strauch E, Takano E, Baylts H, Bibb M. The stringent response in ''Streptomyces coelicolor'' A3(2). ''Molecular Microbiology''. 1991;5(2):289-298.
#Add any other growth factors required for auxotrophic mutants

Latest revision as of 10:25, 19 July 2022

SMM (Supplemented liquid minimal media)

SMM is a 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

  1. With the exception of the trace element solution each solution must be made up and autoclaved separately
  2. The solutions should be mixed in the given order and then made up to 1L with sterile distilled water

References

Strauch E, Takano E, Baylts H, Bibb M. The stringent response in Streptomyces coelicolor A3(2). Molecular Microbiology. 1991;5(2):289-298.

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