CM

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CM (Complete Medium)

Preparation

Per litre:

  • 5g K2HPO4
  • 0.5g NaCl
  • 0.5g MgSO4*7H2O
  • 2g Difco Bacto-peptone
  • 1.5g Difco Casaminoacids
  • 50mg each L-histidine, L-proline, L-tryptophan, and L-tyrosine*
  • 5mL Yeast nucleic acid hydrolysate
  • 1mL vitamin solution
  • 25g glucose
  • 10g agar
  • 1L dH2O

Yeast nucleic acid hydrolysate

  1. Boil 2g yeast nucleic acid in 15mL 1N HCl for 10 min.
  2. Boil 2g yeast nucleic acid in 15mL 1N NaOH for 10 min.
  3. Mix the two solutions and adjust to pH6
  4. Filter through filter paper while hot
  5. Make up the volume to 40mL with water
  6. Do not sterilize

Vitamin solution

  • 100mg riboglavine
  • 100mg nicotinamide
  • 10mg p-aminobenzoic acid
  • 50mg pyridoxine HCl
  • 50mg thiamine HCl
  • 20 mg biotin
  • 100mL water

Instructions

  1. Make up the yeast nucleic acid hydrolysate according to the directions above: do not sterilize. Make up the vitamin solution following the recipe above and sterilize by autoclaving (115˚C, 10 min).
  2. Dissolve all of the ingredients, except the agar and the vitamin solution, but including the glucose, in the distilled water
  3. Adjust the pH to 7.2 with 1N HCl
  4. Add the vitamin solution
  5. Dispense 200ml into flasks each containing 2g agar
  6. Autoclave


Uses

  • A non-selective medium useful for genetic crosses (the progeny from such crosses can subsequently be analysed on appropriate selective media such as MM)[1]

Notes

*The histidine, proline, tryptophan, and tyrosine should be added for the relevant auxotrophic mutants; if not working with auxotrophs, any/all of these amino acids can be omitted.

References

[1] Hopwood DA. Genetic analysis and genome structure in Streptomyces coelicolor. Bacteriol Rev. 1967;31(4):373–403.

[2] Kieser, T., Bibb, M.J., Buttner, M.J., Chater, K.F. and Hopwood, D.A. (2000) Practical Streptomyces Genetics: John Innes Foundation, Norwich Research Park, Colney, Norwich NR4 7UH, UK