CM: Difference between revisions
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(Created page with "=CM (Complete Medium)= ==Preparation== Per litre: *5g K<sub>2</sub>HPO<sub>4</sub> *0.5g NaCl *0.5g MgSO<sub>4</sub>*7H<sub>2</sub>O *2g Difco Bacto-peptone *1.5g Difco Casa...") |
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| Line 9: | Line 9: | ||
*2g Difco Bacto-peptone | *2g Difco Bacto-peptone | ||
*1.5g Difco Casaminoacids | *1.5g Difco Casaminoacids | ||
*50mg <strong>each</strong>L-histidine, L-proline, L-tryptophan, and L-tyrosine<sup>*</sup> | *50mg <strong>each</strong> L-histidine, L-proline, L-tryptophan, and L-tyrosine<sup>*</sup> | ||
*5mL Yeast nucleic acid hydrolysate | *5mL Yeast nucleic acid hydrolysate | ||
*1mL vitamin solution | *1mL vitamin solution | ||
| Line 17: | Line 17: | ||
<strong>Yeast nucleic acid hydrolysate</strong> | <strong>Yeast nucleic acid hydrolysate</strong> | ||
#Boil 2g yeast nucleic acid in 15mL 1N HCl for 10 min. | |||
#Boil 2g yeast nucleic acid in 15mL 1N NaOH for 10 min. | |||
#Mix the two solutions and adjust to pH6 | |||
#Filter through filter paper while hot | |||
#Make up the volume to 40mL with water | |||
#Do not sterilize | |||
<strong>Vitamin solution</strong> | <strong>Vitamin solution</strong> | ||
Revision as of 11:44, 31 July 2019
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
- Boil 2g yeast nucleic acid in 15mL 1N HCl for 10 min.
- Boil 2g yeast nucleic acid in 15mL 1N NaOH for 10 min.
- Mix the two solutions and adjust to pH6
- Filter through filter paper while hot
- Make up the volume to 40mL with water
- 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
- 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).
Dissolve all of the ingredients, except the agar and the vitamin solution, but including the glucose, in the distilled water
- Adjust the pH to 7.2 with 1N HCl
- Add the vitamin solution
- Dispense 200ml into flasks each containing 2g agar
- Autoclave
Uses
Notes
References
Hopwood DA. Genetic analysis and genome structure in Streptomyces coelicolor. Bacteriol Rev. 1967;31(4):373–403.
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
