LisAlgora at 13:27, 4 September 2019

2019-09-04T13:27:30Z

← Older revision		Revision as of 13:27, 4 September 2019
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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 ~~<sup>1</sup>~~Trace element solution		*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

	~~<sup>1</sup>Trace element solution(l<sup>-1</sup>):~~
	**40 mg ZnCl<sub>2</sub>
	**200 mg FeCl<sub>3</sub>.6H<sub>2</sub>O
	**10 mg CuCl<sub>2</sub>.2H<sub>2</sub>O
	**10 mg MnCl<sub>2</sub>.4H<sub>2</sub>O
	**10 mg Na<sub>2</sub>B<sub>4</sub>O<sub>7</sub>.10H<sub>2</sub>O
	**10 mg (NH<sub>4</sub>)<sub>6</sub>Mo<sub>7</sub>O<sub>24</sub>.4H<sub>2</sub>O

LisAlgora: 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 K₂SO₄ *10.12 g..."

2019-07-26T17:16:02Z

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 K2SO4 *10.12 g..."

New page

=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 1Trace element solution
*0.5 ml NaOH (1N) (unsterilised is okay)
*0.75 ml Required growth factors for auxotrophs

1Trace element solution(l-1):
**40 mg ZnCl2
**200 mg FeCl3.6H2O
**10 mg CuCl2.2H2O
**10 mg MnCl2.4H2O
**10 mg Na2B4O7.10H2O
**10 mg (NH4)6Mo7O24.4H2O

==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.

R2YE - Revision history

LisAlgora at 13:27, 4 September 2019

LisAlgora: 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 K2SO4 *10.12 g..."

LisAlgora: 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 K₂SO₄ *10.12 g..."