R2YE: Difference between revisions
(Created page with "=R2YE= R2YE medium is a variation of R2 containing yeast extract. ==Preparation== Make up the following solution: *103 g Sucrose *0.25 g K<sub>2</sub>SO<sub>4</sub> *10.12 g...") |
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*1.5 ml L-proline (20%) | *1.5 ml L-proline (20%) | ||
*10 ml TES buffer (5.73%, adjusted to pH7.2) | *10 ml TES buffer (5.73%, adjusted to pH7.2) | ||
*0.2 ml | *0.2 ml [[Trace element solution]] | ||
*0.5 ml NaOH (1N) (unsterilised is okay) | *0.5 ml NaOH (1N) (unsterilised is okay) | ||
*0.75 ml Required growth factors for auxotrophs | *0.75 ml Required growth factors for auxotrophs | ||
Latest revision as of 13:27, 4 September 2019
R2YE
R2YE medium is a variation of R2 containing yeast extract.
Preparation
Make up the following solution:
- 103 g Sucrose
- 0.25 g K2SO4
- 10.12 g MgCl2.6H2O
- 10 g Glucose
- 0.1 g Difco Casaminoacids
- 800 ml dH2O
Pour 80 ml of the solution into 250 ml Erlenmeyer flasks each containing 2.2 g Difco Bacto agar. Sterilise by autoclaving. At the time of use, remelt the medium and add to each flask the following autoclaved solutions in the order listed:
- 5 ml Difco yeast extract (10%)
- 1 ml KH2PO4 (0.5%)
- 8 ml CaCl2.2H2O (3.68%)
- 1.5 ml L-proline (20%)
- 10 ml TES buffer (5.73%, adjusted to pH7.2)
- 0.2 ml Trace element solution
- 0.5 ml NaOH (1N) (unsterilised is okay)
- 0.75 ml Required growth factors for auxotrophs
Uses
- Protoplasts regeneration
Notes
Streptomyces clavuligerus does not utilise glucose and it must be substituted by glycerol.
References
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
Thompson CJ, Ward JM, Hopwood DA (1980) DNA cloning in Streptomyces: resistance genes from antibiotic-producing species. Nature, 286, 525-527.
