| Size | Price | Stock | Qty |
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| 50mg |
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| 100mg |
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| 250mg |
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| Other Sizes |
| ln Vitro |
5-Fluoroorotic acid at 1 mg/ml in minimal medium (Table I) completely inhibits the growth of all Ura+ yeast strains tested, while Ura- cells can grow and form resistant colonies. [1]
When used at a lower concentration (500 μg/ml), background growth of Ura+ cells is considerably greater. [1] On YPD rich medium containing 1 mg/ml 5-Fluoroorotic acid, the growth inhibition is variable: some Ura+ strains are prevented from growing while others are not. [1] |
|---|---|
| Cell Assay |
Yeast cells are pre-grown in liquid YPD medium or on YPD plates before selection on 5-Fluoroorotic acid medium. Up to 10⁷ cells can be plated on a single 50 mm Petri dish containing 5-FOA medium. Resistant colonies appear within 4–7 days at 30°C. [1]
For allele replacement (transplacement), Ura+ transformants containing an integrated plasmid with URA3 and the gene of interest are plated onto medium containing 5-Fluoroorotic acid. Cells that lose the URA3 gene via homologous recombination become 5-FOA resistant (Ura-), while parental Ura+ cells do not grow. [1] In plasmid shuffling, a strain carrying a chromosomal deletion of an essential gene (cdc27Δ1) and a wild-type copy on a URA3 plasmid is transformed with a mutagenized LEU2 plasmid carrying the same gene. Leu+ transformants are grown at permissive temperature, then replica-plated onto 5-Fluoroorotic acid medium. Ura- papillae that grow out of the background are analyzed. If the LEU2 plasmid carries a temperature-sensitive allele, papillae appear only at permissive temperature (22°C) but not at restrictive temperature (36°C). If the LEU2 plasmid carries a null mutation, no 5-FOA-resistant papillae appear at any temperature. [1] To introduce ura3 deletions, yeast cells transformed with integrating plasmid pJEF1332 (containing ura3Δ1 mutation and TRP1 marker) are selected on SC-Trp medium. Transformants are then streaked onto 5-Fluoroorotic acid medium (containing tryptophan and other nutrients) to isolate 5-FOA-resistant segregants that have lost the wild-type URA3 gene, plasmid sequences, and TRP1. [1] |
| Toxicity/Toxicokinetics |
5-Fluoroorotic acid at 1 mg/ml in minimal medium is toxic to all Ura+ yeast strains tested, blocking their growth. [1]
On YPD rich medium containing 1 mg/ml 5-Fluoroorotic acid, toxicity is variable: it prevents growth of some Ura+ strains but not others. [1] The compound is not toxic to Ura- cells (e.g., ura3 mutants), allowing their selective growth. [1] |
| References | |
| Additional Infomation |
5-Fluoroorotic acid (5-FOA) is a selective agent that allows detection of rare ura3- cells among a large population of wild-type Ura+ cells. Its utility stems from the availability of URA3-based cloning vectors, the small size and known sequence of the URA3 gene, and the ease and efficacy of the selection. [1]
The selection works by conversion of the nontoxic compound to one that is toxic to wild-type Ura+ cells; mutant cells lacking the ability to form the toxic compound grow in the presence of the inert precursor. [1] 5-FOA selection is used in transplacement (allele replacement) to replace a chromosomal wild-type gene with an in vitro-generated mutant allele, and in plasmid shuffling to isolate conditional lethal mutations in essential cloned genes. [1] It is also used to measure mitotic recombination frequencies of yeast duplications: the sequence of interest is cloned into Yip5 and integrated, then the frequency of Ura- segregants (5-FOA-resistant colonies) is measured. [1] The complete deletion of the URA3 gene (1.1 kb HindIII fragment) is lethal to the cell, suggesting an essential gene lies 5’ to URA3. The ura3-52 mutation is nonreverting but contains all wild-type URA3 sequences due to Ty element insertion. [1] |
| Molecular Formula |
C5H3FN2O4
|
|---|---|
| Molecular Weight |
174.0867
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| Exact Mass |
174.007
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| CAS # |
703-95-7
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| PubChem CID |
69711
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| Appearance |
White to yellow solid powder
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| Density |
1.8±0.1 g/cm3
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| Melting Point |
278 °C (dec.)(lit.)
|
| Index of Refraction |
1.580
|
| LogP |
-1
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
1
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| Heavy Atom Count |
12
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| Complexity |
309
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| Defined Atom Stereocenter Count |
0
|
| InChi Key |
SEHFUALWMUWDKS-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C5H3FN2O4/c6-1-2(4(10)11)7-5(12)8-3(1)9/h(H,10,11)(H2,7,8,9,12)
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| Chemical Name |
5-fluoro-2,4-dioxo-1H-pyrimidine-6-carboxylic acid
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ~125 mg/mL (~718.02 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (11.95 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.08 mg/mL (11.95 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 2.08 mg/mL (11.95 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 5.7442 mL | 28.7208 mL | 57.4416 mL | |
| 5 mM | 1.1488 mL | 5.7442 mL | 11.4883 mL | |
| 10 mM | 0.5744 mL | 2.8721 mL | 5.7442 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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