R5: Difference between revisions
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(Created page with "=R5= ==Preparation== Make up the following solution: *103 g Sucrose *0.25 g K<sub>2</sub>SO<sub>4</sub> *10.12 g MgCl<sub>2</sub>.6H<sub>2</sub>O *10 g Glucose *0.1 g Difco...") |
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*10 g Glucose | *10 g Glucose | ||
*0.1 g Difco Casaminoacids | *0.1 g Difco Casaminoacids | ||
*2 ml [[trace | *2 ml [[trace element solution]] | ||
*5 g Difco yeast extract | *5 g Difco yeast extract | ||
*5.73 g TES buffer | *5.73 g TES buffer | ||
Revision as of 15:12, 26 September 2019
R5
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
- 2 ml trace element solution
- 5 g Difco yeast extract
- 5.73 g TES buffer
- To 1000 ml dH2O
Pour 100 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:
- 1 ml KH2PO4 (0.5%)
- 0.4 ml CaCl2.2H2O (5M, 3.68%)
- 1.5 ml L-proline (20%)
- 0.7 ml NaOH (1N) (unsterilised is okay)
- 0.75 ml Required growth factors for auxotrophs
Uses
- See R2YE - just an alternate way of making but folklore suggests less reproducible results on R5.
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
