Freimoser:Recipes:Yeast:SC Sfree: Difference between revisions
(New page: ===Sulfur free SC-medium (self mixed SC medium)=== =Remark:= This recipe describes the mixing of a sulfur free SC-medium. All sulfur sources in “normal” SC-medium are replaced by oth...) |
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=Remark:= | =Sulfur free SC-medium (self mixed SC medium)= | ||
===Remark:=== | |||
This recipe describes the mixing of a sulfur free SC-medium. All sulfur sources in “normal” SC-medium are replaced by other salts, mostly chlorides. The changes are also indicated. | This recipe describes the mixing of a sulfur free SC-medium. All sulfur sources in “normal” SC-medium are replaced by other salts, mostly chlorides. The changes are also indicated. | ||
The phosphate source is separate in order to allow mixing of media with different phosphate content. | The phosphate source is separate in order to allow mixing of media with different phosphate content. | ||
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The described procedure works, but it is possible that there are simpler solutions. But it is not possible to make a stock solution of all trace elements for example. | The described procedure works, but it is possible that there are simpler solutions. But it is not possible to make a stock solution of all trace elements for example. | ||
=20x Salt solution, 500 ml= | ===20x Salt solution, 500 ml=== | ||
MgCl<sub>2</sub>: 16 g (4 mM) | MgCl<sub>2</sub>: 16 g (4 mM) | ||
NaCl: 1 g (1.7 mM) | NaCl: 1 g (1.7 mM) | ||
CaCl<sub>2</sub> x 2 H<sub>2</sub>O: 1 g (1 mM) | CaCl<sub>2</sub> x 2 H<sub>2</sub>O: 1 g (1 mM) | ||
=100 x phosphate source, 100 ml= | ===100 x phosphate source, 100 ml=== | ||
KH<sub>2</sub>PO<sub>4</sub>: 10 g (7.5 mM) | |||
=1000x trace element solution 1, 10 ml= | ===1000x trace element solution 1, 10 ml=== | ||
Boric acid (H<sub>3</sub>BO<sub>3</sub>): 0.05 g (8 uM) | Boric acid (H<sub>3</sub>BO<sub>3</sub>): 0.05 g (8 uM) | ||
CuCl<sub>2</sub> x 2 H<sub>2</sub>O: 0.004 g (0.25 uM) (replaces CuSO<sub>4</sub>) | CuCl<sub>2</sub> x 2 H<sub>2</sub>O: 0.004 g (0.25 uM) (replaces CuSO<sub>4</sub>) | ||
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=MnCl<sub>2</sub> x 4 H<sub>2</sub>O (manganese chloride):= 0.04 g in 10 ml (1000x) (2 uM) (replaces MnSO<sub>4</sub>) | =MnCl<sub>2</sub> x 4 H<sub>2</sub>O (manganese chloride):= 0.04 g in 10 ml (1000x) (2 uM) (replaces MnSO<sub>4</sub>) | ||
=Na<sub>2</sub>MoO<sub>4</sub> (sodium molybdate):= 0.02 g in 10 ml (1000x) (1 uM) | ===Na<sub>2</sub>MoO<sub>4</sub> (sodium molybdate):=== 0.02 g in 10 ml (1000x) (1 uM) | ||
=FeCl<sub>3</sub> x 6 H<sub>2</sub>O (ferric chloride):= 0.02 g in 10 ml (500x) (1.5 uM) | ===FeCl<sub>3</sub> x 6 H<sub>2</sub>O (ferric chloride):=== 0.02 g in 10 ml (500x) (1.5 uM) | ||
NH<sub>4</sub>Cl (ammonium chloride): 2 g in 10 ml (100x, 37.8 mM final conc. In medium) | NH<sub>4</sub>Cl (ammonium chloride): 2 g in 10 ml (100x, 37.8 mM final conc. In medium) | ||
Revision as of 02:52, 1 February 2007
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Sulfur free SC-medium (self mixed SC medium)
Remark:
This recipe describes the mixing of a sulfur free SC-medium. All sulfur sources in “normal” SC-medium are replaced by other salts, mostly chlorides. The changes are also indicated. The phosphate source is separate in order to allow mixing of media with different phosphate content. The 20x salt solution, 100x phosphate source and 40% glucose solution were autoclaved. All other solutions were filter sterilized. The described procedure works, but it is possible that there are simpler solutions. But it is not possible to make a stock solution of all trace elements for example.
20x Salt solution, 500 ml
MgCl2: 16 g (4 mM) NaCl: 1 g (1.7 mM) CaCl2 x 2 H2O: 1 g (1 mM)
100 x phosphate source, 100 ml
KH2PO4: 10 g (7.5 mM)
1000x trace element solution 1, 10 ml
Boric acid (H3BO3): 0.05 g (8 uM) CuCl2 x 2 H2O: 0.004 g (0.25 uM) (replaces CuSO4) KI (potassium iodide): 0.01 g (0.6 uM) C4H10O6Zn (zinc acetate): 0.055 g (2 uM) (replaces ZnSO4)
Separate solutions:
=MnCl2 x 4 H2O (manganese chloride):= 0.04 g in 10 ml (1000x) (2 uM) (replaces MnSO4)
===Na2MoO4 (sodium molybdate):=== 0.02 g in 10 ml (1000x) (1 uM)
===FeCl3 x 6 H2O (ferric chloride):=== 0.02 g in 10 ml (500x) (1.5 uM)
NH4Cl (ammonium chloride): 2 g in 10 ml (100x, 37.8 mM final conc. In medium)
Additional components:
=Glucose (C6H12O6):= 40% (20x)
=MES buffer (2-(N-morpholino)ethanesulfonic acid):= 0.5 M, pH 6 (10x)
YNB 1000x vitamin solution
