MM: Difference between revisions
(Created page with "=MM Medium= ==Preparation== Per litre: *0.5g L-asparagine *0.5g K2HPO4 *0.2g MgSO4*7H2O *0.01g FeSO4*7H2O *10g glucose (added after autoclaving) *10g agar *=1L dH2O '''Instr...") |
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=MM Medium= | =MM (Minimal Medium)= | ||
MM is a minimal medium suitable for growth of many <em>Streptomyces</em> sp. It is often made up with glucose or mannitol, but other carbon sources can be used. It can be further supplemented with additional growth factors as described below. | |||
==Preparation== | ==Preparation== | ||
| Line 20: | Line 21: | ||
==Uses== | ==Uses== | ||
*studies of auxotrophic mutants and carbon source utilization | |||
==Notes== | ==Notes== | ||
The glucose can be replaced by alternative carbon sources such as glycerol, arabinose, or mannitol (each at 5g/L). Mannitol is often preferable to glucose for optimal growth and development. | |||
To study carbon source utilization, the L-asparagine should be replaced by ammonium sulfate (5 g/L(NH4)2SO4) - to avoid confusion due to utilization of asparagine as a carbon source. | |||
Additional supplements can be added as follows (amount/L): amino acids except histidine, 50 mg; histidine, 70 mg; adenine and uracil, 10 mg; vitamins, 1 mg; dihydrostreptomycin sulfate, 50 mg; acriflavine, 15 mg. (Hopwood 1967) | |||
==References== | ==References== | ||
Hopwood DA. Genetic analysis and genome structure in Streptomyces coelicolor. Bacteriol Rev. 1967;31(4):373–403. | 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 | |||
Revision as of 13:21, 26 June 2019
MM (Minimal Medium)
MM is a minimal medium suitable for growth of many Streptomyces sp. It is often made up with glucose or mannitol, but other carbon sources can be used. It can be further supplemented with additional growth factors as described below.
Preparation
Per litre:
- 0.5g L-asparagine
- 0.5g K2HPO4
- 0.2g MgSO4*7H2O
- 0.01g FeSO4*7H2O
- 10g glucose (added after autoclaving)
- 10g agar
- =1L dH2O
Instructions
- Dissolve all of the ingredients except the agar and the glucose in the distilled water
- Adjust the pH to 7.0-7.2
- Dispense 200ml into flasks each containing 2g agar
- Autoclave
- After autoclaving, add 4 ml sterile 50% glucose solution to each flask
- Add any other growth factors required for auxotrophic mutants
Uses
- studies of auxotrophic mutants and carbon source utilization
Notes
The glucose can be replaced by alternative carbon sources such as glycerol, arabinose, or mannitol (each at 5g/L). Mannitol is often preferable to glucose for optimal growth and development.
To study carbon source utilization, the L-asparagine should be replaced by ammonium sulfate (5 g/L(NH4)2SO4) - to avoid confusion due to utilization of asparagine as a carbon source.
Additional supplements can be added as follows (amount/L): amino acids except histidine, 50 mg; histidine, 70 mg; adenine and uracil, 10 mg; vitamins, 1 mg; dihydrostreptomycin sulfate, 50 mg; acriflavine, 15 mg. (Hopwood 1967)
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
