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FAM-xtra™ Phosphoramidite

There are several ways of labeling an oligonucleotide with fluorescein. 6-FAM phosphoramidite might be the most popular fluorescent building block that is widely used to efficiently label an oligonucleotide at the 5’-end with the fluorescein fluorophore. Standard cleavage and deprotection with ammonium hydroxide are used to liberate the fluorescein-labeled oligos. Although 6-FAM has be ubiquitously used for developing a variety of qPCR probes, its fluorescence suffers two serve limitations: a). highly pH dependent; and b). photo-bleach rapidly. FAM-xtra™ Phosphoramidite has been developed to address these two limitations while the well-beloved operational convenience of 6-FAM phosphoramidite is intactly maintained. FAM-xtra Phosphoramidite can be used exactly as is used for 6-FAM phosphoramidite under the same operation conditions without any changes needed. The oligos made from FAM-xtra Phosphoramidite have quite a few great benefits: a). FAM-xtra-labeled oligos have the identical spectra to those of 6-FAM labeled ones; b). FAM-xtra-labeled oligos are much brighter than the corresponding FAM-labeled ones; c). FAM-xtra-labeled oligos have much higher photostability than the corresponding FAM-labeled ones; and d). The fluorescence of FAM-xtra-labeled oligos is much less pH sensitive than that of the corresponding FAM-labeled ones. Under the physiological conditions, the fluorescence of FAM-xtra-labeled oligos is essentially pH-independent, making the FAM-xtra-labeled oligos much more robust qPCR probes.

Spectrum

Product family

NameExcitation (nm)Emission (nm)Extinction coefficient (cm -1 M -1)Correction Factor (260 nm)Correction Factor (280 nm)
FAM-xtra CPG *500A*493517830000.320.178
FAM-xtra CPG *1000A*493517830000.320.178
6-FAM phosphoramidite [5'-Fluorescein phosphoramidite] *CAS 204697-37-0*493517830000.320.178

References

View all 8 references: Citation Explorer
Development of an Innovative Intradermal siRNA Delivery System Using a Combination of a Functional Stearylated Cytoplasm-Responsive Peptide and a Tight Junction-Opening Peptide.
Authors: Ibaraki, Hisako and Kanazawa, Takanori and Takashima, Yuuki and Okada, Hiroaki and Seta, Yasuo
Journal: Molecules (Basel, Switzerland) (2016)
Non-nucleoside phosphoramidites of xanthene dyes (FAM, JOE, and TAMRA) for oligonucleotide labeling.
Authors: Kvach, Maksim V and Tsybulsky, Dmitry A and Shmanai, Vadim V and Prokhorenko, Igor A and Stepanova, Irina A and Korshun, Vladimir A
Journal: Current protocols in nucleic acid chemistry (2013): Unit 4.55
Practical synthesis of isomerically pure 5- and 6-carboxytetramethylrhodamines, useful dyes for DNA probes.
Authors: Kvach, Maksim V and Stepanova, Irina A and Prokhorenko, Igor A and Stupak, Aleksander P and Bolibrukh, Dmitry A and Korshun, Vladimir A and Shmanai, Vadim V
Journal: Bioconjugate chemistry (2009): 1673-82
Synthesis and cellular uptake of fluorescently labeled multivalent hyaluronan disaccharide conjugates of oligonucleotide phosphorothioates.
Authors: Karskela, Marika and Virta, Pasi and Malinen, Melina and Urtti, Arto and Lönnberg, Harri
Journal: Bioconjugate chemistry (2008): 2549-58
Synthesis of HyBeacons and dual-labelled probes containing 2'-fluorescent groups for use in genetic analysis.
Authors: Dobson, Neil and McDowell, David G and French, David J and Brown, Lynda J and Mellor, John M and Brown, Tom
Journal: Chemical communications (Cambridge, England) (2003): 1234-5
Page updated on October 9, 2024

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Physical properties

Molecular weight

879.94

Solvent

MeCN

Spectral properties

Correction Factor (260 nm)

0.32

Correction Factor (280 nm)

0.178

Extinction coefficient (cm -1 M -1)

83000

Excitation (nm)

493

Emission (nm)

517

Storage, safety and handling

H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22

Storage

Freeze (< -15 °C); Minimize light exposure
UNSPSC12171501