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Specifications
| Appearance: | Off White to Pale Yellow Powder |
| Assay (31P NMR): | 95% min |
| Assay (HPLC): | 95% min |
| Water: | 0.4% max |
Applications
1. Fluorescent Labeling of Oligonucleotides
Used in solid-phase synthesis to incorporate fluorescein directly at the 5'-end of DNA or RNA strands.
Enables creation of fluorescently labeled probes, primers, and molecular beacons for various applications.
2. Molecular Biology and Diagnostics
Fluorescein-labeled oligonucleotides serve as probes in fluorescence in situ hybridization (FISH), real-time PCR (qPCR), and microarray assays.
Used for sensitive detection of nucleic acid sequences in research and diagnostic contexts.
3. Fluorescence Resonance Energy Transfer (FRET) Probes
Incorporated into oligonucleotides to design FRET-based sensors and probes for studying molecular interactions and conformational changes.
4. Nucleic Acid Interaction Studies
Facilitates monitoring of hybridization events, binding affinities, and enzyme activities involving nucleic acids.
Benefits
1. Site-Specific Incorporation
Enables precise placement of fluorescein at the 5'-end during synthesis for consistent and reproducible labeling.
2. High Fluorescence Efficiency
Fluorescein provides bright green fluorescence with excellent quantum yield, enhancing sensitivity in detection assays.
3. Compatibility with Automated Synthesis
Designed for use in standard oligonucleotide synthesizers, streamlining probe production.
4. Versatility
Suitable for various oligonucleotide lengths and sequences.
Can be combined with other modifications for multiplexed assays.
5. Stable and Reliable
Provides robust fluorescence with minimal photobleaching during experiments.
Conclusion
5-Fluorescein phosphoramidite (CAS 204697-37-0) is a specialized reagent used to introduce fluorescein fluorescent labels at the 5'-end of synthetic oligonucleotides via automated DNA/RNA synthesis. This modification is essential for producing fluorescent probes used in molecular diagnostics, gene expression analysis, FISH, and nucleic acid interaction studies. Its compatibility with solid-phase synthesis and bright fluorescence make it a valuable tool in biotechnology and molecular biology research.
