6-FAM-phosphoramidite
Structural formula | ||||||||||
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General | ||||||||||
Surname | 6-FAM-phosphoramidite | |||||||||
other names |
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Molecular formula | C 46 H 58 N 3 O 10 P | |||||||||
Brief description |
pale yellow powder |
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properties | ||||||||||
Molar mass | 843.94 g mol −1 | |||||||||
Physical state |
firmly |
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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 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 .
Individual evidence
- ↑ a b c data sheet 6-FAM-phosphoramidite from Sigma-Aldrich , accessed on May 22, 2017 ( PDF ).
Web links
- Thomas A. Schild: Introduction to Real-Time Taqman PCR Technology . 7700 SDS workshop