Difference between revisions of "5-Fluoroorotic Acid (5-FOA)"
From PombEvolution
Jump to navigationJump to searchm |
|||
| Line 1: | Line 1: | ||
5-FOA (C<sub>5</sub>H<sub>3</sub>FN<sub>2</sub>O) counter selects for ura+ strains. The nontoxic 5-FOA is converted to toxic 5-flurouracil in yeast with a functional [[Ura4 selection marker|ura4]] gene, which normally synthesizes uracil. We can thus select for strains that are ura-.<BR> | 5-FOA (C<sub>5</sub>H<sub>3</sub>FN<sub>2</sub>O) counter selects for ura+ strains. The nontoxic 5-FOA is converted to toxic 5-flurouracil in yeast with a functional [[Ura4 selection marker|ura4]] gene, which normally synthesizes uracil. We can thus select for strains that are ura-.<BR> | ||
* Make a 100X stock solution of 5-FOA in DMSO by adding 100 mg/ml. <BR> | * Make a 100X stock solution of 5-FOA in DMSO by adding 100 mg/ml. <BR> | ||
| − | * For use, use 1:100 (200 µl/plate).<BR> | + | * For use, use 1:100 (200 µl/9cm plate or 3µl/300µl in Deep Well Plate).<BR> |
* Stock solution is available in freezer 7.<BR> | * Stock solution is available in freezer 7.<BR> | ||
* Keep plates in fridge. | * Keep plates in fridge. | ||
Revision as of 12:34, 14 April 2022
5-FOA (C5H3FN2O) counter selects for ura+ strains. The nontoxic 5-FOA is converted to toxic 5-flurouracil in yeast with a functional ura4 gene, which normally synthesizes uracil. We can thus select for strains that are ura-.
- Make a 100X stock solution of 5-FOA in DMSO by adding 100 mg/ml.
- For use, use 1:100 (200 µl/9cm plate or 3µl/300µl in Deep Well Plate).
- Stock solution is available in freezer 7.
- Keep plates in fridge.
For a good protocol on how to use this with YES, see: Petersen & Russell
