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6-Fluorescein phosphoramidite

There are several ways of labeling an oligonucleotide with fluorescein. The choice of label is diversified further, depending on the spectral requirements. 5'-fluorescein-CE phosphoramidite, derived from the single isomer 6-carboxyfluorescein, 5'-hexachlorofluorescein-CE phosphoramidite (HEX) and 5'-tetrachlorofluorescein-CE phosphoramidite (TET) can all be used to efficiently label an oligonucleotide at the 5'-end. Labeling the 3'-end of an oligo with fluorescein can be achieved using 3'-Fluorescein CPGs. Standard cleavage and deprotection with ammonium hydroxide liberates the fluorescein-labeled oligo when using any of these supports.
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Physical properties
Molecular weight1176.35
SolventMeCN
Spectral properties
Absorbance (nm)487
Correction Factor (260 nm)0.32
Correction Factor (280 nm)0.35
Extinction coefficient (cm -1 M -1)800001
Excitation (nm)498
Emission (nm)517
Quantum yield0.79001, 0.952
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure
UNSPSC12171501
CAS204697-37-0
Calculators

Common stock solution preparation

Table 1. Volume of MeCN needed to reconstitute specific mass of 6-Fluorescein phosphoramidite to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

0.1 mg0.5 mg1 mg5 mg10 mg
1 mM85.009 µL425.044 µL850.087 µL4.25 mL8.501 mL
5 mM17.002 µL85.009 µL170.017 µL850.087 µL1.7 mL
10 mM8.501 µL42.504 µL85.009 µL425.044 µL850.087 µL

Molarity calculator

Enter any two values (mass, volume, concentration) to calculate the third.

Mass (Calculate)Molecular weightVolume (Calculate)Concentration (Calculate)Moles
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Spectrum
References
View all 13 references: Citation Explorer
Sequence-specific, self-reporting hairpin inversion probes
Authors: Browne KA., undefined
Journal: J Am Chem Soc (2005): 1989
Modified oligonucleotides containing lithocholic acid in their backbones: their enhanced cellular uptake and their mimicking of hairpin structures
Authors: Kim SJ, Bang EK, Kwon HJ, Shim JS, Kim BH.
Journal: Chembiochem (2004): 1517
Synthesis of HyBeacons and dual-labelled probes containing 2'-fluorescent groups for use in genetic analysis
Authors: Dobson N, McDowell DG, French DJ, Brown LJ, Mellor JM, Brown T.
Journal: Chem Commun (Camb) (2003): 1234
Reduced aggregation and improved specificity of G-rich oligodeoxyribonucleotides containing pyrazolo[3,4-d]pyrimidine guanine bases
Authors: Kutyavin IV, Lokhov SG, Afonina IA, Dempcy R, Gall AA, Gorn VV, Lukhtanov E, Metcalf M, Mills A, Reed MW, S and ers S, Shishkina I, Vermeulen NM.
Journal: Nucleic Acids Res (2002): 4952
Diels--Alder bioconjugation of diene-modified oligonucleotides
Authors: Hill KW, Taunton-Rigby J, Carter JD, Kropp E, Vagle K, Pieken W, McGee DP, Husar GM, Leuck M, Anziano DJ, Sebesta DP.
Journal: J Org Chem (2001): 5352