Inquiry Basket
Fluorescein-dT phosphoramidite
| Catalog Number | A19-0088 |
| Category | DNA Stains |
| Molecular Formula | C79H87N6O17P |
| Molecular Weight | 1423.54 |
| Catalog Number | Size | Price | Quantity |
|---|---|---|---|
| A19-0088 | -- | $-- |
- Worldwide Delivery
- Quality Assurance
- 24/7 Customer Service
Product Introduction
This phosphoramidite allows fluorescein (FAM) to be introduced into any position in the oligonucleotide sequence during synthesis by the phosphoramidite method (in the middle of the chain, at the 5’- and
3’-terminus). The reagent is a conjugate of deoxythymidine phosphoramidite
and 6-isomer of FAM. Modification is performed during oligonucleotide synthesis by substituting standard dT phosphoramidite with fluorescein-dT phosphoramidite. This modification does not affect exonuclease or polymerase activity.,For modification with fluorescein at the 5’-terminus use
FAM phosphoramidite, 6-isomer.,Coupling: 10 minutes,Deprotection: standard conditions using ammonium hydroxide;
deprotection time depends on oligonucleotide composition and nucleobase
protecting groups (deprotection for 17 h at 55 °С removes all protecting
groups from standard nucleobases). AMA (solution of 30% ammonium
hydroxide/40% aqueous methylamine 1:1 v/v) can be used with ~5% of
non-fluorescent side product forming. To avoid formation of the side product, start deprotection with ammonium hydroxide (30 min at room temperature), then add an equal volume of 40% aqueous methylamine and continue deprotection as required with AMA (e.g. 10 min at 65 °C).
Chemical Information |
|
|---|---|
| Purity | NMR 1H, NMR 31P, HPLC-MS |
| Solubility | Good in acetonitrile, DCM |
| Appearance | Off White powder |
- Product Specification
- Application
| CF260 | 0.20 |
| CF280 | 0.17 |
| Fluorescence Quantum Yield | 0.93 |
| Excitation | 490 |
| Emission | 513 |
| Storage | Storage: 12 months after receival at -20 °C in the Dark. Transportation: at room temperature for up to 3 weeks. Avoid prolonged exposure to light. Desiccate. |
Fluorescein-dT phosphoramidite, a fluorescent nucleotide analog utilized in diverse molecular biology and genetic research endeavors, boasts a multitude of applications. Here are key applications of fluorescein-dT phosphoramidite creatively:
Fluorescent DNA Probes: A staple in the realm of genetic exploration, fluorescein-dT phosphoramidite serves as a linchpin in crafting fluorescently labeled DNA probes for hybridization assays. These meticulously engineered probes exhibit a remarkable affinity for specific DNA sequences, enabling their visualization and detection in cutting-edge techniques like FISH (Fluorescence In Situ Hybridization). This advancement revolutionizes the study of chromosome rearrangements and gene localization across a myriad of organisms.
Real-Time PCR: In the thrilling domain of real-time PCR assays, the incorporation of fluorescein-dT phosphoramidite into primer or probe sequences heralds a new era of DNA amplification detection. The fluorescent signal emitted during the PCR process correlates directly with the quantity of synthesized DNA, facilitating meticulous quantitative analysis. This breakthrough proves pivotal in pivotal applications such as gene expression studies, pathogen detection, and genotyping endeavors.
DNA Sequencing: A cornerstone in the field of genetic decoding, fluorescein-dT phosphoramidite shines brightly in Sanger sequencing as a premier fluorescent label for dideoxynucleotide terminators. This ingenious application sees the incorporation of the labeled nucleotide, generating fragments of varying lengths which are subsequently resolved through capillary electrophoresis. The captured fluorescence signals pave the way for accurate determination of DNA sequences, propelling genetic research to new heights of precision.
Applications of Fluorescent Probes & Dyes
Get More Product Details?
