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| Product Name | 5′-FAM (Fluorescein) Labeled Oligonucleotide |
| Catalog No. | PRIPRO-0005 |
| Description | A custom DNA oligonucleotide with a 5′-terminal 6-carboxyfluorescein (6-FAM) fluorescent label attached via a six-carbon amino linker during solid-phase synthesis. FAM is the most widely used green fluorophore with excitation maximum at 495 nm and emission at 520 nm, making it compatible with the FAM/SYBR Green channel on all major qPCR instruments and capillary electrophoresis sequencers. The oligo is HPLC purified to remove unincorporated dye and truncated sequences, ensuring a single fluorescent species. Supplied as a dried pellet quantified by UV-Vis spectrophotometry with both A260 and dye absorbance measurements reported on the certificate of analysis. |
| Intended Use | Designed for use as a fluorescent primer in fragment analysis, microsatellite genotyping, fluorescent PCR, DNA sequencing with capillary electrophoresis, and as a labeled probe for fluorescence in situ hybridization (FISH). |
| Principle / Technology | Solid-phase phosphoramidite synthesis with 6-FAM phosphoramidite coupling at the 5′ terminus via aminohexyl linker |
| Detection Method | Fluorescence spectroscopy (Ex 495 nm / Em 520 nm); HPLC with dual UV and fluorescence detection |
| Sample Type | PCR amplicons, genomic DNA for genotyping, chromosomal preparations for FISH |
| Performance Range / Specifications | 10–60 bases standard; dye attachment verified by absorbance at 495 nm |
| Sensitivity / LOD | Fluorescence detection limit typically 0.1–1 nM in standard capillary electrophoresis instruments |
| Specificity | Sequence-specific hybridization; dye does not interfere with base pairing |
| Reaction Conditions / Protocol | Resuspend in nuclease-free water or TE buffer to 100 µM stock. Protect from light at all times. Use 0.1–0.5 µM final concentration in PCR with detection in FAM channel. |
| Components / Formulation | Single tube containing lyophilized FAM-labeled oligonucleotide |
| Storage Conditions | –20 °C in the dark; minimize light exposure during handling |
| Shelf Life | 12 months at –20 °C dry; 6 months at –20 °C in solution (protected from light) |
| Package Specifications | Amber or foil-wrapped microcentrifuge tube; 5–50 nmol scale options |
| Product Form | Lyophilized pellet (typically pale yellow to orange from dye) |
| Quality Control | HPLC purity ≥90%; MALDI-TOF MS mass confirmation; UV-Vis absorbance at 260 nm and 495 nm with dye incorporation efficiency reported; fluorescence emission spectrum verified. |
| Key Features | Bright green fluorescence with high quantum yield; compatible with all major capillary sequencers and qPCR platforms; stable 6-carbon linker minimizes dye quenching; HPLC purified; dye/coupling efficiency reported on COA. |
| Purity | ≥90% by HPLC (dual-wavelength detection); single dye-labeled species |
| Concentration | Lyophilized; resuspend to 100 µM stock based on provided OD260 and calculated extinction coefficient |
| Activity / Unit Definition | Fluorescence quantum yield ~0.9; extinction coefficient ~75,000 M⁻¹cm⁻¹ at 495 nm |
| Molecular Weight | Calculated from oligo sequence plus ~537 Da for 6-FAM linker moiety |
| Source / Origin | Synthetic oligonucleotide with chemically coupled fluorophore |
| pH Range / Optimal pH | Optimal fluorescence at pH 7.5–9.0; FAM fluorescence is pH-sensitive and decreases below pH 7.0 |
| Shipping Conditions | Ambient temperature; lyophilized dye-labeled oligos shipped in light-protective packaging |
| Expiration Date / Stability | 12 months dry at –20 °C in the dark; 6 months in TE buffer at –20 °C protected from light |
| Regulatory / Compliance | For research use only; not for diagnostic procedures |
| Compatibility | FAM channel (Ex ~495 nm / Em ~520 nm) on ABI PRISM, Bio-Rad CFX, Roche LightCycler, Qiagen Rotor-Gene, and other qPCR platforms |
| Recommended Buffer System | TE buffer pH 8.0 or nuclease-free water; avoid Tris buffers below pH 7.0 for fluorescence measurement |
| Application Notes / Precautions | Protect from light during storage and handling. For capillary electrophoresis, dilute to 1–10 nM in Hi-Di formamide with size standard. Verify fluorescence before use by brief UV illumination. |
| Batch-to-Batch Consistency | Dye incorporation efficiency >90% by HPLC peak area; fluorescence intensity per OD260 unit consistent within ±10% across batches |
For research use only, not for clinical use.
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