|Molecular formula||C 46 H 58 N 3 O 10 P|
pale yellow powder
|External identifiers / databases|
|Molar mass||843.94 g mol −1|
|As far as possible and customary, SI units are used. Unless otherwise noted, the data given apply to standard conditions .|
6-FAM-phosphoramidite , also 6-FAM-amidite , is the phosphoramidite derivative of 6-carboxyfluorescein (FAM), a fluorescent dye that is used in the synthesis of oligonucleotides to generate hybridization probes for real-time PCR assays and thus is used for the quantitative detection of nucleic acid sequences .
6-FAM-phosphoramidite is a reporter dye that is covalently bound to the 5 'end of the probe, which is responsible for the color signal of the reaction. The excitation wavelength of 6-FAM-phosphoramidite is 488 nm, the emission wavelength 518 nm. A quencher dye is bound to the 3 'end of the probe (e.g. TAMRA). The Taq polymerase equipped with a 5 ' exonuclease activity removes the reporter dye from the DNA . After the complete synthesis of the PCR product, the quencher dye is also released. Due to the spatial separation, the reporter dye emits a fluorescence signal after excitation. If, on the other hand, the intact probe is excited, the quencher dye suppresses its fluorescence due to the spatial proximity to the reporter dye .
- Thomas A. Schild: Introduction to Real-Time Taqman PCR Technology . 7700 SDS workshop