6-FAM-phosphoramidit - 6-FAM-phosphoramidit
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|Brief description|| |
pale yellow powder 
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|Molar mass||843.94 g · mol −1|
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|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 at the 3 'end of the probe (e.g. TAMRA). The Taq polymerase equipped with a 5'- exonuclease activity dissolves the reporter dye from the DNAfrom. After the PCR product has been completely synthesized, 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