SMM: Difference between revisions

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(Created page with "=SMM (Supplemented liquid minimal media)= Strauch et al., 1991<sup>1</sup> ==Preparation== Per litre: *81.9 mL PEG 6000 (BDH 6.1% w/v in distilled water) *2.5 mL Mg<sub>2</su...")
 
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*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 bugger (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
#After autoclaving, add 4 ml sterile 50% glucose solution (autoclaved separately or filter-sterilized) to each flask
#Add any other growth factors required for auxotrophic mutants

Revision as of 15:42, 1 August 2019

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
  • 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
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