5-Fluoroorotic acid

Catalog No.S9873 Batch:S987301

Technical Data

Formula	C₅H₃FN₂O₄
Molecular Weight	174.09	CAS No.	703-95-7
Solubility (25°C)*	In vitro	DMSO	35 mg/mL (201.04 mM)
		Water	Insoluble
		Ethanol	Insoluble
* <1 mg/ml means slightly soluble or insoluble. * Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations. * Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

Preparing Stock Solutions

Biological Activity

Description	5-Fluoroorotic acid (5-FOA, 5-Fluoroorotate, NSC 31712) is a selective agent in yeast molecular genetics. 5-Fluoroorotic acid possesses a well-expressed anticandidal effect close to that of 5-fluorocytosine, as well as moderate antidermatophytal effects.
In vitro	5-FOA is an extremely useful reagent for the selection of Ura- cells amid a population of Ura+ cells. The selection is effective in transformation and recombination studies where loss of URA3+ is desired. A new plasmid shuffling procedure based on the 5-FOA^R selection permits the recovery of conditional lethal mutations in cloned genes that encode vital functions.^[1]

Protocol (from reference)

Cell Assay:^{^[1]}	Cell lines Yeast cells Concentrations 1 mg/ml, 500 μg/ml Incubation Time 4-7 days Method Yeast cells are generally pregrown in liquid YPD medium or on YPD plates prior to selection on 5-FOA. As many as 10⁷ cells may be plated on a single, 50 mm Petri dish containing 5-FOA medium. Resistant colonies will grow up within 4-7 days at 30°. The most useful concentration of the drug is generally 1 mg/ml 5-fluoroorotic acid, but in the interest of economy it is possible to use as little as 500 μg/ml 5-FOA. When lower concentrations of 5-FOA are used, however, background growth of the Ura+ cells is considerably greater. This background growth is less of a problem when Ura- derivatives occur at a high frequency (e.g., when a URA3+ marker is located between the elements of a direct duplication). In some instances it may be desirable to use a rich medium such as YPD as opposed to the minimal medmm normally used with 5-FOA selection.

Cell Assay:^{^[1]}

Cell lines
Yeast cells
Concentrations
1 mg/ml, 500 μg/ml
Incubation Time
4-7 days
Method
Yeast cells are generally pregrown in liquid YPD medium or on YPD plates prior to selection on 5-FOA. As many as 10⁷ cells may be plated on a single, 50 mm Petri dish containing 5-FOA medium. Resistant colonies will grow up within 4-7 days at 30°. The most useful concentration of the drug is generally 1 mg/ml 5-fluoroorotic acid, but in the interest of economy it is possible to use as little as 500 μg/ml 5-FOA. When lower concentrations of 5-FOA are used, however, background growth of the Ura+ cells is considerably greater. This background growth is less of a problem when Ura- derivatives occur at a high frequency (e.g., when a URA3+ marker is located between the elements of a direct duplication). In some instances it may be desirable to use a rich medium such as YPD as opposed to the minimal medmm normally used with 5-FOA selection.

References

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SHIPPING AND STORAGE
Selleck products are transported at room temperature. If you receive the product at room temperature, please rest assured, the Selleck Quality Inspection Department has conducted experiments to verify that the normal temperature placement of one month will not affect the biological activity of powder products. After collecting, please store the product according to the requirements described in the datasheet. Most Selleck products are stable under the recommended conditions.

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